Abstract Breast density is a significant independent risk factor for breast cancer; women with dense breasts have a 4-6-fold increased risk of the disease compared to women with non-dense breasts. It has been suggested that 30% of all breast cancer cases occur in women with > 50% dense areas. Breast density reflects variations in breast tissue composition. It is characterized by high proportions of stroma, containing collagen and other ECM proteins, fibroblasts, endothelial cells, and immune cells, suggesting a pro-tumor inflammatory microenvironment. To investigate molecular differences between dense and nondense breast tissue and their association with triple-negative breast cancer (TNBC), we used the PanCancer IO 360 panel to explore RNA expression and signaling pathway regulation across the samples. We observed differences in RNA expression among subjects, thereby identifying distinct molecular groups in dense breasts. The subjects were clustered into 3 subtypes with distinct molecular signatures and biological pathways. We identified a group (G1) with increased expression of genes involved in inflammatory processes, but still maintains a closer association with the phenotype of non-dense breasts. We also identified two other groups (G2 and G3) that are more distinct regarding the non-dense group and different from each other. These groups are related to reorganization and cellular architecture in the microenvironment (G2) and to proliferative regulation, cancer pathways, and increased gene expression in tumor development (G3). These groups do not seem to correlate with age or menopause stage in this first screen; instead, they resemble more subtypes of density or stages of density development. Additionally, analyzing RNA expression in TNBC samples, we identify an association with dense G3 samples that express more RNAs related to DNA repair, with upregulation of pathways in DNA damage, epigenetic regulation, apoptosis, and metabolic stress. At the same time, the pathways such as interferon signaling, immune cell adhesion and migration, and NF-kappa-beta are downregulated. These results suggest that specific density subtypes with distinct molecular signatures may promote tumor development more than others. To better understand this association and determine whether there are similarities in the resident cells of dense breast and TNBC samples, we are conducting spatial biology analyses to explore their microenvironments. The accurate classification of these subtypes has the potential to impact breast cancer prevention strategies and early diagnosis. Further investigations with a larger cohort are underway to understand better how molecular alterations in dense groups contribute to tumor development. Citation Format: Jay William Fox, Natalia Dworak, Patcharin Pramoonjago, Christopher A. Moskaluk, Ana Karina de Oliveira. Distinct molecular signatures in dense breast tissue as potential drivers of breast cancer development [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 6318.

Angioimmunoblastic T-cell lymphoma (AITL) and follicular T-cell lymphoma (FTCL) represent the two most well-defined manifestations of follicular helper T-cell lymphoma. The overlapping clinical, immunophenotypic and genetic features of these T follicular helper (TFH) cell-derived neoplasms belie their divergent growth patterns while suggesting that both are part of the same disease spectrum. Recent single-cell transcriptomic analyses provide a comprehensive description of the tumor microenvironment (TME) in AITL, with functional studies further indicating a role for non-malignant populations in AITL pathobiology. Meanwhile, the composition and role of the TME in FTCL remains understudied. In particular, the use of emerging spatial transcriptomics technologies has rarely been applied to follicular helper T-cell lymphomas of any variety. To better asses the TME composition in FTCL, we performed spatial transcriptomics using the GeoMx® Digital Spatial Profiler (NanoString Technologies) on nodal follicular helper T-cell lymphomas of follicular type (n = 3 tumors) and angioimmunoblastic type (n = 4 tumors) as well as on benign lymphoid follicles from sections of reactive tonsil (n = 4). Regions of interest (ROI; 4 per tumor) were identified through the evaluation of H&E-stained sections as well as routine diagnostic immunohistochemical stains. A trio of morphology markers consisting of a pan-T-cell antigen (CD3) and a pair of sensitive TFH antigens (PD-1 and ICOS) was used to define populations of TFH-like cells for microdissection, while the remaining tissue within each respective ROI was designated as bulk TME. Compared to their corresponding bulk TME, the TFH-like fractions from both the FTCL and AITL groups exhibited significantly greater expression of genes corresponding to the employed morphology markers (CD3D, CD3E, CD3G, PDCD1, ICOS) as well as genes encoding additional TFH markers (CXCR5 and/or CXCL13) and numerous other TFH-related genes (e.g., BATF, CD40LG, IL21, MAF, TIAM1, TCF7, TNFRSF4, TOX, TOX2), supporting the validity of this approach. Upon dimensional reduction, the bulk TME from the nodular FTCL follicles formed an extended cluster either adjacent to (UMAP) or partially within (tSNE) the bulk TME from tumor cell-rich regions of AITL, while both groups were separate from the microenvironment of benign lymphoid follicles. Quantification of immune and non-immune cell subtypes was performed using SpatialDecon. Compared to benign lymphoid follicles, the follicles of FTCL were significantly enriched in macrophages, memory CD8+ T cells, NK cells, plasmacytoid dendritic cells and myeloid dendritic cells; however, only memory CD8+ T cells and myeloid dendritic cells remained significant after accounting for the intrinsically B-cell-rich nature of the benign follicles. FTCL follicles were proportionally enriched in naïve B cells compared to tumor cell-rich regions of AITL, in keeping with the presence of mantle zone-type lymphocytes in both the follicular lymphoma-like and progressive transformation of germinal centers-like growth patterns of FTCL. No other statistically significant differences in immune cell subtype composition were observed between FTCL and AITL in this small cohort. In conclusion, this work represents the successful application of spatial transcriptomics to follicular helper T-cell lymphomas, and is the first such study to include FTCL. Our findings highlight significant differences in the composition of FTCL follicles compared to benign lymphoid follicles as well as substantial overlap between the cellular compositions of FTCL and AITL, in keeping with the plethora of shared biology between these entities. Further analysis will benefit from technologies affording single-cell resolution.

The long-term physiological consequences of respiratory viral infections, particularly in the aftermath of the COVID-19 pandemic-termed post-acute sequelae of SARS-CoV-2 (PASC)-are rapidly evolving into a major public health concern1-3. While the cellular and molecular aetiologies of these sequelae are poorly defined, increasing evidence implicates abnormal immune responses3-6 and/or impaired organ recovery7-9 after infection. However, the precise mechanisms that link these processes in the context of PASC remain unclear. Here, with insights from three cohorts of patients with respiratory PASC, we established a mouse model of post-viral lung disease and identified an aberrant immune-epithelial progenitor niche unique to fibroproliferation in respiratory PASC. Using spatial transcriptomics and imaging, we found a central role for lung-resident CD8+ T cell-macrophage interactions in impairing alveolar regeneration and driving fibrotic sequelae after acute viral pneumonia. Specifically, IFNγ and TNF derived from CD8+ T cells stimulated local macrophages to chronically release IL-1β, resulting in the long-term maintenance of dysplastic epithelial progenitors and lung fibrosis. Notably, therapeutic neutralization of IFNγ + TNF or IL-1β markedly improved alveolar regeneration and pulmonary function. In contrast to other approaches, which require early intervention10, we highlight therapeutic strategies to rescue fibrotic disease after the resolution of acute disease, addressing a current unmet need in the clinical management of PASC and post-viral disease.

The radiosensitization potential of focused ultrasound (FUS)-induced mild hyperthermia was assessed in an allogenic subcutaneous C6 glioma tumor model in rats. Mild hyperthermia at 42 °C was induced in tumors using a single-element 350 kHz FUS transducer. Radiation was delivered with a small animal radiation research platform using a single-beam irradiation technique. The combined treatment involved 20 min of FUS hyperthermia immediately before radiation. Tumor growth changes were observed one week post-treatment. A radiation dose of 2 Gy alone showed limited tumor control (30% reduction). However, when combined with FUS hyperthermia, there was a significant reduction in tumor growth compared to other treatments (tumor volumes: control—1174 ± 554 mm3, FUS-HT—1483 ± 702 mm3, 2 Gy—609 ± 300 mm3, FUS-HT + 2 Gy—259 ± 186 mm3; ANOVA p < 0.00001). Immunohistological analysis suggested increased DNA damage as a short-term mechanism for tumor control in the combined treatment. In conclusion, FUS-induced mild hyperthermia can enhance the effectiveness of radiation in a glioma tumor model, potentially improving the outcome of standard radiation treatments for better tumor control.

Lymphadenopathy is a common finding in patients with IgG4-related disease (IgG4-RD) and often associated with increased IgG4+ plasma cells in this setting. The histologic features of so-called IgG4-related lymphadenopathy (IgG4-LAD) have seldom been investigated in children and adolescents, and step-wise progression to extranodal IgG4-RD has not been described. This study was performed to further evaluate the frequency, pathologic features, and clinical significance of IgG4-LAD-like histologic changes in the pediatric setting. We analyzed 37 benign lymph nodes collected semi-consecutively from children aged 0–18 years at our institution for both absolute and relative IgG4+ plasma cell abundance and recurrent histomorphologic patterns associated with IgG4-LAD. The combination of IgG4+/IgG+ plasma cell ratio >40

Pathological and inflammatory events in muscle after the injection of snake venoms vary in different regions of the affected tissue and at different time intervals. In order to study such heterogeneity in the immune cell microenvironment, a murine model of muscle necrosis based on the injection of the venom of Daboia russelii was used. Histological and immunohistochemical methods were utilized to identify areas in muscle tissue with a different extent of muscle cell damage, based on the presence of hypercontracted muscle cells, a landmark of necrosis, and on the immunostaining for desmin. A gradient of inflammatory cells (neutrophils and macrophages) was observed from heavily necrotic areas to less damaged and non-necrotic areas. GeoMx® Digital Spatial Profiler (NanoString, Seattle, WA, USA) was used for assessing the presence of markers of various immune cells by comparing high-desmin (nondamaged) and low-desmin (damaged) regions of muscle. Markers of monocytes, macrophages, M2 macrophages, dendritic cells, neutrophils, leukocyte adhesion and migration markers, and hematopoietic precursor cells showed higher levels in low-desmin regions, especially in samples collected 24 hr after venom injection, whereas several markers of lymphocytes did not. Moreover, apoptosis (BAD) and extracellular matrix (fibronectin) markers were also increased in low-desmin regions. Our findings reveal a hitherto-unknown picture of immune cell microheterogeneity in venom-injected muscle which greatly depends on the extent of muscle cell damage and the time lapse after venom injection.

Cancer cell dissemination to sentinel lymph nodes is associated with poor patient outcomes, particularly in breast cancer. The process by which cancer cells egress from the primary tumor upon interfacing with the lymphatic vasculature is complex and driven by dynamic interactions between cancer cells and stromal cells, including cancer-associated fibroblasts (CAF). The matricellular protein periostin can distinguish CAF subtypes in breast cancer and is associated with increased desmoplasia and disease recurrence in patients. However, as periostin is secreted, periostin-expressing CAFs are difficult to characterize in situ, limiting our understanding of their specific contribution to cancer progression. Here, we used in vivo genetic labeling and ablation to lineage trace periostin+ cells and characterize their functions during tumor growth and metastasis. Periostin-expressing CAFs were spatially found at periductal and perivascular margins, were enriched at lymphatic vessel peripheries, and were differentially activated by highly metastatic cancer cells versus poorly metastatic counterparts. Surprisingly, genetically depleting periostin+ CAFs slightly accelerated primary tumor growth but impaired intratumoral collagen organization and inhibited lymphatic, but not lung, metastases. Periostin ablation in CAFs impaired their ability to deposit aligned collagen matrices and inhibited cancer cell invasion through collagen and across lymphatic endothelial cell monolayers. Thus, highly metastatic cancer cells mobilize periostin-expressing CAFs in the primary tumor site that promote collagen remodeling and collective cell invasion within lymphatic vessels and ultimately to sentinel lymph nodes. Significance: Highly metastatic breast cancer cells activate a population of periostin-expressing CAFs that remodel the extracellular matrix to promote escape of cancer cells into lymphatic vessels and drive colonization of proximal lymph nodes.

The long-term physiological consequences of SARS-CoV-2, termed Post-Acute Sequelae of COVID-19 (PASC), are rapidly evolving into a major public health concern. The underlying cellular and molecular etiology remain poorly defined but growing evidence links PASC to abnormal immune responses and/or poor organ recovery post-infection. Yet, the precise mechanisms driving non-resolving inflammation and impaired tissue repair in the context of PASC remain unclear. With insights from three independent clinical cohorts of PASC patients with abnormal lung function and/or viral infection-mediated pulmonary fibrosis, we established a clinically relevant mouse model of post-viral lung sequelae to investigate the pathophysiology of respiratory PASC. By employing a combination of spatial transcriptomics and imaging, we identified dysregulated proximal interactions between immune cells and epithelial progenitors unique to the fibroproliferation in respiratory PASC but not acute COVID-19 or idiopathic pulmonary fibrosis (IPF). Specifically, we found a central role for lung-resident CD8+ T cell-macrophage interactions in maintaining Krt8hi transitional and ectopic Krt5+ basal cell progenitors, thus impairing alveolar regeneration and driving fibrotic sequelae after acute viral pneumonia. Mechanistically, CD8+ T cell derived IFN-γ and TNF stimulated lung macrophages to chronically release IL-1β, resulting in the abnormal accumulation of dysplastic epithelial progenitors and fibrosis. Notably, therapeutic neutralization of IFN-γ and TNF, or IL-1β after the resolution of acute infection resulted in markedly improved alveolar regeneration and restoration of pulmonary function. Together, our findings implicate a dysregulated immune-epithelial progenitor niche in driving respiratory PASC. Moreover, in contrast to other approaches requiring early intervention, we highlight therapeutic strategies to rescue fibrotic disease in the aftermath of respiratory viral infections, addressing the current unmet need in the clinical management of PASC and post-viral disease.

AbstractCancer cell dissemination to sentinel lymph nodes is associated with poor patient outcomes, particularly in breast cancer. The process by which cancer cells egress from the primary tumor upon interfacing with the lymphatic vasculature is complex and driven by dynamic interactions between cancer cells and stromal cells, including cancer-associated fibroblasts (CAF). The matricellular protein periostin can distinguish CAF subtypes in breast cancer and is associated with increased desmoplasia and disease recurrence in patients. However, as periostin is secreted, periostin-expressing CAFs are difficult to characterize in situ, limiting our understanding of their specific contribution to cancer progression. Here, we used in vivo genetic labeling and ablation to lineage trace periostin+ cells and characterize their functions during tumor growth and metastasis. Periostin-expressing CAFs were spatially found at periductal and perivascular margins, were enriched at lymphatic vessel peripheries, and were differentially activated by highly metastatic cancer cells versus poorly metastatic counterparts. Surprisingly, genetically depleting periostin+ CAFs slightly accelerated primary tumor growth but impaired intratumoral collagen organization and inhibited lymphatic, but not lung, metastases. Periostin ablation in CAFs impaired their ability to deposit aligned collagen matrices and inhibited cancer cell invasion through collagen and across lymphatic endothelial cell monolayers. Thus, highly metastatic cancer cells mobilize periostin-expressing CAFs in the primary tumor site that promote collagen remodeling and collective cell invasion within lymphatic vessels and ultimately to sentinel lymph nodes.Significance:Highly metastatic breast cancer cells activate a population of periostin-expressing CAFs that remodel the extracellular matrix to promote escape of cancer cells into lymphatic vessels and drive colonization of proximal lymph nodes.

Breast cancer affects over 2,000,000 women worldwide, with more than 680,000 deaths per year. Increased breast density is one of the risk factors for breast cancer, albeit less than the risk associated with increased age and genetic mutation. It has been suggested that high breast density accounts for 15% of the breast cancers diagnosed. On radiographic examination, breast density appears as opaque regions associated with increased cellularity and matrix, which in some circumstances contribute to difficulty observing small tumors. Dense regions in the breast have been associated with both morphological changes as well as molecular changes in the tissue. How these changes may play a role in increased risk for tumorigenesis are not well described. In this study, we examine the partial spatial transcriptomes of specific regions in normal breast tissue containing dense regions, including regions of normal density, the interface of dense and non-dense regions and the dense regions themselves using the GeoMx Digital Spatial Profiling system (DSP). Multi-label immunofluorescence was used to distinguish tissue morphology in FFPE samples from two health patients. We selected 24 regions of interest (ROIs) from each tissue section included dense, interface and non-dense breast. Using barcoded DNA oligos attached in situ hybridization probes (for RNA) via ultraviolet (UV)- photocleavable linkers, we screened over 1800 genes from Cancer Transcriptome Atlas panel. Our results indicated an elevated expression of CD68 and CD33, markers for macrophages and myeloid-derived suppressor cells (MDSCs) in dense breast regions as compared to the non-dense areas. Identifying markers for myeloid lineage cells was followed by an increased expression of the genes EOMES, TIGIT and RORA, which are engaged in the immunosuppressive phenotype. This could be hypothesized to support a pro-tumorigenesis microenvironment preventing T cells from recognizing newly arisen tumor cells. In addition, we also observed a significant expression of cytokines (IL12A and IL26) and chemokines (CCL19 and CXCL10) in the dense breast, which are involved in the regulation of inflammatory response and chemotaxis of monocytes and T-lymphocytes cells. Both CCL19 and IL26 have been demonstrated to play a role in cell proliferation. Thus, these initial studies suggest that the dense breast microenvironment is potentially a pro-tumorigenic environment that, with sufficient factors, gives rise to tumorigenesis. These data also suggest potential nodes of therapeutic intervention that may lower such risk. Citation Format: Ana Karina de Oliveira, Patcharin Pramoonjago, Christopher A. Moskaluk, Jay W. Fox. Spatial transcriptomic evidence for an inflammatory, pro tumorigenic microenvironment in normal human dense breast tissue [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2023.

AbstractObjectiveDevelop a deep learning-based methodology using the foundations of systems pathology to generate highly accurate predictive tools for complex gastrointestinal diseases, using celiac disease (CD) as a prototype.DesignTo predict the severity of CD, defined by Marsh–Oberhüber classification, we used deep learning to develop a model based on histopathologic features.ResultsThe study was based on a pediatric cohort of 124 patients identified with different classes of CD severity. The model predicted CD with an overall 88.7% accuracy with the highest for Marsh IIIc (91.0%; 95% sensitivity; 91% specificity). The model identified EECs as a defining feature of children with Marsh IIIc CD and endocrinopathies which was confirmed using immunohistochemistry.ConclusionThis deep learning image analysis platform has broad applications in disease treatment, management, and prognostication and paves the way for precision medicine.SummaryWhat is already known about this subject?–Deep Learning has the potential to generate predictive models for complex gastrointestinal diseases.What are the new findings?–Our deep learning-based model used the foundations of systems pathology to generate a highly accurate predictive tool for complex gastrointestinal diseases, using a celiac disease (CD) pediatric cohort as a prototype.–The model predicated CD severity with high accuracy and identified enteroendocrine cells as a defining feature of children with severe CD and endocrinopathies.How might it impact on clinical practice in the foreseeable future?–Assessment of histopathological markers at the time of diagnosis that can predict risk of severity or complications can have broad applications in disease treatment, management, and prognostication and pave the way for precision medicine.

Bone marrow (BM) lymphocyte subsets are evaluated by flow cytometry or immunohistochemistry for diagnostic purposes; however, CD4:CD8 T lymphocyte ratios are often erroneously interpreted using peripheral blood ranges. There are few and no recent studies describing the composition of lymphocytes within the marrow space, or normal reference ranges. Lymphocyte subsets in cytopenic patients and hospital autopsy BM specimens were evaluated to better characterize CD4:CD8 ratios. Ten patients with a history of cytopenia were identified from 2017 to 2021. Clinical history, cytogenetic testing, and results of a next generation sequencing panel were reviewed to rule out hematolymphoid disease. Thirty-five decedents who underwent a hospital autopsy from 2018 to 2019 were identified. History of hematolymphoid disease was ruled out by chart review. Immunohistochemical staining for CD3, CD20, CD4, and CD8 was evaluated with digital image analysis. Findings were compared to peripheral blood flow cytometry in a group of 20 living patients. BM CD4:CD8 ratios by image analysis were significantly lower than peripheral blood, mean in cytopenic patients 0.37:1 and mean in decedents 0.51:1 versus 2.6:1 ( p = < .001 in both groups). BM CD4:CD8 ratios were significantly lower ( p = 0.04) than ratios found using flow cytometry on the same specimen, suggesting hemodilution. There was no significant difference in CD4:CD8 ratios when comparing living patients and decedents’ marrows ( p = > 0.99). Lymphoid aggregates were encountered with increasing frequency in older individuals. These findings aid in the evaluation of BM lymphocyte subsets and distribution both in living patients and autopsy evaluation. We also present a practical approach to image analysis.

ABSTRACT:Basal cell carcinoma (BCC) is the most common skin cancer, and it has numerous histologic mimics with variable prognoses and treatments. Although some immunohistochemical stains can be used for the differential diagnosis of BCC, variability and overlap in results can complicate their interpretation. Immunohistochemical staining for glioma-associated oncogene-1 (Gli-1) was performed on 26 nodular BCCs, 22 infiltrative BCCs, 9 basaloid squamous cell carcinomas, 12 desmoplastic trichoepitheliomas, 19 Merkel cell carcinomas, 11 sebaceous carcinomas, 10 cylindromas, 14 spiradenomas, 12 adenoid cystic carcinomas (AdCC), and 1 solitary trichoepithelioma. Strength of staining was scored as 0, 1+, 2+, or 3+, and distribution of staining was categorized as diffuse, multifocal, or focal. Strong, diffuse Gli-1 expression was seen in all tumors with basal epidermal-type differentiation, including BCC, trichoepithelioma, and basaloid squamous cell carcinoma. All examples of Merkel cell carcinoma were negative for cytoplasmic expression. Seven out of 11 sebaceous carcinomas were negative for Gli-1, and the remaining 4 showed 1+ expression. Cylindroma, spiradenoma, and AdCC, each an adnexal skin tumor, showed the most variable staining, but with cylindroma and spiradenoma demonstrating comparable labeling patterns. Overall, although Gli-1 may not distinguish between basal epidermal-type tumors, it may have a role in separating that group from lesions with adnexal differentiation, particularly sebaceous carcinoma, but also cylindroma, spiradenoma, and AdCC. Any cytoplasmic staining seems to exclude the diagnosis of Merkel cell carcinoma.

Meiotic arrest is a common cause of human male infertility, but the causes of this arrest are poorly understood. Transactive response DNA-binding protein of 43 kDa (TDP-43) is highly expressed in spermatocytes in the preleptotene and pachytene stages of meiosis. TDP-43 is linked to several human neuro-degenerative disorders wherein its nuclear clearance accompanied by cytoplasmic aggregates underlies neurodegeneration. Exploring the functional requirement for TDP-43 for spermatogenesis for the first time, we show here that conditional KO (cKO) of the Tardbp gene (encoding TDP-43) in male germ cells of mice leads to reduced testis size, depletion of germ cells, vacuole formation within the seminiferous epithelium, and reduced sperm production. Fertility trials also indicated severe subfertility. Spermatocytes of cKO mice showed failure to complete prophase I of meiosis with arrest at the midpachytene stage. Staining of synaptonemal complex protein 3 and gamma H2AX, markers of the meiotic synaptonemal complex and DNA damage, respectively, and super illumination microscopy revealed nonhomologous pairing and synapsis defects. Quantitative RT-PCR showed reduction in the expression of genes critical for prophase I of meiosis, including Spo11 (initiator of meiotic double-stranded breaks), Rec8 (meiotic recombination protein), and Rad21L (RAD21-like, cohesin complex component), as well as those involved in the retinoic acid pathway critical for entry into meiosis. RNA-Seq showed 1036 upregulated and 1638 downregulated genes (false discovery rate <0.05) in the Tardbp cKO testis, impacting meiosis pathways. Our work reveals a crucial role for TDP-43 in male meiosis and suggests that some forms of meiotic arrest seen in infertile men may result from the loss of function of TDP-43.

Clostridium difficile (C. difficile) incidence has tripled over the past 15 years and is attributed to the emergence of hypervirulent strains. While it is clear that C. difficile toxins cause damaging colonic inflammation, the immune mechanisms protecting from tissue damage require further investigation. Through a transcriptome analysis, we identify IL-33 as an immune target upregulated in response to hypervirulent C. difficile. We demonstrate that IL-33 prevents C. difficile-associated mortality and epithelial disruption independently of bacterial burden or toxin expression. IL-33 drives colonic group 2 innate lymphoid cell (ILC2) activation during infection and IL-33 activated ILC2s are sufficient to prevent disease. Furthermore, intestinal IL-33 expression is regulated by the microbiota as fecal microbiota transplantation (FMT) rescues antibiotic-associated depletion of IL-33. Lastly, dysregulated IL-33 signaling via the decoy receptor, sST2, predicts C. difficile-associated mortality in human patients. Thus, IL-33 signaling to ILC2s is an important mechanism of defense from C. difficile colitis.

We introduce here recently developed highly resolved Sub-Terahertz resonance spectroscopy of biological molecules and cells combined with molecular dynamics (MD) computational analysis as a new approach for optical visualization and quantification of the presence of microRNAs, particularly the mir-200 family, as potential biomarkers in samples from tissue of epithelial ovarian cancers for disease early detection, analysis, prognosis and treatment.METHOD: A set of samples for this study was prepared from anonymized archival formalin-fixed, paraffin-embedded ovarian epithelial tissue containing regions of invasive neoplastic cells from cases of high-histologic grade serous papillary ovarian carcinoma. Control samples were normal mucosa from fallopian tubes of patients with no known malignancy. Spectroscopic characterization of tissue samples in this study was performed using a continuous wave, frequency domain automated spectrometer operating at room temperature in the spectral region of 310-500 GHz. The spectral results were compared with molecular dynamics simulations and absorption coefficient calculations utilized to predict the absorption spectra.RESULTS: The characteristic spectroscopic features in absorption spectra, particularly the presence of absorption peaks near 13 cm-1 have been identified as cancer indicators. Tissue samples heterogeneity, reflected by diverse spectral signatures, provides additional, very specific information that may be used for identification of cancer subtypes, clinical behavior or sensitivity to specific therapies. Further work is warranted to determine if this signature can be detected in bio-fluids from ovarian cancer patients. If strongly correlated with cancer burden, it may then be investigated as a potential new biomarker for disease monitoring, and also perhaps as a biomarker for cancer screening.