Goal: Emerging evidence in diverse tumor types establishes a link between lymphatic dissemination and collective tumor cell invasion. To simulate the biomechanical features of the tumor-lymphatic microenvironment, we developed a 3D tumor-lymphatic architecture biomimetic (T-LAB) platform. Methods: Mathematical and computational fluid dynamics modeling were used to determine the fluid flow, oscillatory flow-induced shear stress, and system pressure in the 3D-printed macrofluidics platform. Results: Various human breast cancer cell lines and human dermal lymphatic endothelial cells (HDLEC) were seeded on a matrix in the T-LAB and imaged for up to 96h to assess cell morphology, viability, migration, and invasion. Co-culture of inflammatory breast cancer cells with HDLEC in the T-LAB, determined to simulate the fluidic properties of the tumor lymphatic microenvironment, demonstrated tumor cell clusters/emboli formation and collective invasion similar to the clinicopathological features observed in patients. Conclusions: The 3D T-LAB model developed here can be used to culture any type of tumor cell to study topographical features that impact tumor-lymphatic interface, collective invasion, and lymphatic dissemination.

Abstract Background Patient and Public Involvement (PPI) is increasingly recognised as essential in research design and funding. While benefits for patients include empowerment and trust in science, PPI also offers unique learning opportunities for early-stage researchers (ESRs), however, structured experiential PPI training for ESRs remains limited. The objective of this work is to report on the impact of a structured PPI engagement event, Be the Voice in Your Community of Health Research on ESRs’ confidence, skills, and understanding of PPI, while supporting patients from diverse ethnic backgrounds to share their lived experiences in cancer care and research. Methods A one-day workshop held in 2025 at University College Dublin (UCD) combined short talks, patient panels and small-group discussions with facilitation. Each facilitation group included one senior researcher, who co-facilitated the session alongside the ESRs. The senior researcher provided oversight, supported discussion flow, and ensured methodological consistency, while the ESRs led participant engagement and interactive components. Participants comprised 5 senior researchers, 10 early-stage researchers (ESRs; PhD students and research assistants working in cancer biology, molecular oncology, bioinformatics and computational biology), and 20 community participants from minority ethnic backgrounds living in Ireland, including three individuals with lived experience of cancer. Post-event reflections and an anonymised questionnaire captured qualitative and quantitative feedback from the ESRs. Descriptive statistics summarised Likert-type items. Qualitative data were thematically analysed. Results Nine of 10 ESRs rated the event ‘excellent’ ( one ‘good’ ). Self-rated confidence in speaking with patients increased (mean of 4.3 before to 6.0 after; 7-point scale). Qualitative themes indicated (i) development of communication, (ii) active listening and empathy (iii) renewed research motivation and (iv) intention to adopt more inclusive research practices. ESRs showed a desire for more frequent training opportunities of this nature. Community participants reported feeling heard and respected and emphasised the importance of cultural awareness and interpersonal respect. Conclusions This event demonstrated the dual value of PPI by creating a structured yet informal setting that fostered open dialogue and mutual learning, empowering patients while enriching researchers’ understanding and practice. Embedding structured PPI training early in research careers may help cultivate a generation of empathetic, socially responsible scientists and advance more inclusive, patient-centred cancer research.

Inflammatory breast cancer (IBC) is an aggressive breast cancer subtype characterized by tumor emboli, lymphovascular invasion (LVI), and early dissemination. Herein, we establish adaptive stress response (ASR) as a biologic feature linking stress adaptation to tumor emboli survival, lymphatic dissemination, therapeutic response, and disparities. Using a previously defined 226 ASR-related genes, complementary preclinical models of tumor emboli and lymphatic circulating cell clusters, and independent patient cohorts, we identified ASR genes enriched for XIAP-NFκB, oxidative stress response, inflammatory, and immune pathways. CXCL8 emerged as one of the most highly upregulated transcripts in tumor emboli and was shared across both models; however, CXCL8 , IL6 , and PTGS2 were downregulated in lymphatic circulating cell clusters and LVI-positive triple-negative IBC patients, suggesting dynamic remodeling of inflammatory signaling during dissemination. CYP4B1 was associated with ER status, LVI, and therapeutic response across multiple cohorts, implicating metabolic stress adaptation in dissemination. IL6 and PTGS2 were elevated in self-reported Black patients with triple-negative IBC compared to White patients. Pharmacologic inhibition of XIAP-NFκB and oxidative stress pathways suppressed tumor emboli formation. Collectively, these findings identify ASR signaling as a framework linking tumor emboli survival, dissemination, and therapeutic vulnerability in IBC.

Inflammatory breast cancer (IBC) is a rare yet highly aggressive subtype of breast cancer, characterized by distinct clinical features, rapid progression, and complex biology. Despite decades of research, patient outcomes remain poor, underscoring the need for innovative approaches. The International IBC Symposium 2025 convened global experts to discuss advances in the field and to outline strategic priorities for research and clinical care. This report summarizes the key themes discussed. A major topic was IBC biology, with particular emphasis on tumor emboli formation and tumor microenvironment interactions, which shape the distinctive characteristics of the disease. Updates in molecular and immune profiling were presented, illustrating how these data may guide personalized therapeutic approaches. From a therapeutic standpoint, the importance of trimodal therapy in patients with nonmetastatic IBC was reinforced, while highlighting the need to investigate novel agents, rational drug combinations, and biologically informed strategies to improve outcomes. Additional sessions addressed optimization of locoregional management, even for selected patients with metastatic disease. Experts also discussed emerging technologies, including spatial transcriptomics, artificial intelligence tools, and liquid biopsy, as promising methods to advance disease characterization, facilitate earlier detection, and refine disease monitoring. Finally, a common theme across presentations was the critical importance of expanding awareness and education, as well as fostering global collaboration through the IBC International Consortium to standardize diagnosis, accelerate translational research, and ensure equitable access to care. Collectively, these efforts aim to redefine the clinical trajectory of IBC and move the field closer to achieving durable disease control and, ultimately, curative outcomes.

Inflammatory breast cancer (IBC) is a rare but aggressive subtype of breast cancer characterized by rapid progression and poor prognosis. Despite its distinct clinical presentation and molecular features, the immune landscape of IBC and its potential role in driving the aggressive phenotype remain poorly understood. This study aimed to characterize the spatial immune landscape of IBC, compare it with that of subtype-matched non-inflammatory breast cancer (nIBC), and evaluate the prognostic implications of immune cell composition and localization. We analyzed pre-treatment tumor samples from 161 IBC and 115 subtype-matched nIBC patients using immunohistochemistry (IHC) for CD8, FOXP3, CD79α, CD163, and PD-L1. Digital image analysis quantified the immune cell density and relative marker area in the tumor area (TA) and invasive margin (IM). Associations with clinicopathological features, pathological response to neoadjuvant chemotherapy (NACT), and survival were assessed using multivariate logistic regression and Cox proportional hazards models. Transcriptomic validation was performed using Affymetrix gene expression data and consensus TME deconvolution. IBC showed higher infiltration of CD163 + tumor-associated macrophages (TAMs) compared to nIBC. Gene expression data confirmed IHC findings, and pathway analysis linked high TAM density with inflammatory and proliferative pathways. The spatial distribution of immune cells was prognostically relevant, with high CD8 + T-cell infiltration (OR: 0.41, 95

PURPOSE:Inflammatory breast cancers (IBC) are characterized by diffuse clusters of cells found in dermal lymphatic vessels, known as tumor emboli. IBC needs a novel treatment because it is the most aggressive breast cancer form. We hypothesized that the interaction between tumor emboli and the tumor immune microenvironment (TiME) fosters survival signaling, leading to the aggressiveness of disease. METHODS:Ex vivo tumor emboli were generated from patient-derived cell lines cultured in a lymphatic-like platform and subjected to transcriptomic and proteomic analysis. Spatial immunophenotyping was performed on IBC patient samples with tumor emboli. A transgenic CX3cr1GFP murine model was generated for visualization of macrophages and tumor emboli within the TiME via a surgically implanted window chamber, enabling intravital imaging and targeting. RESULTS:Gene and protein analyses of tumor emboli cultures compared to 2D monolayer cultures revealed upregulation of TNFR signaling networks, CXCL8, and immune cell chemotaxis genes. Spatial immunophenotyping of tumor emboli in patient samples demonstrated high levels of CD163+ tumor-associated macrophages. Furthermore, intravital imaging of CX3cr1GFP mice confirmed macrophage movement toward tumor cell clusters. Finally, targeting macrophage-associated TNF-α-signaling using SMAC mimetic, Birinapant, inhibited the tumor emboli phenotype in vivo. CONCLUSIONS:This study, the first to our knowledge, identifies TNF-α signaling and macrophage infiltration in IBC tumor emboli. Therapeutic strategies targeting TNF-α signaling to induce cell death and reduced macrophage influence have the potential to improve IBC outcomes.

Disclosure: G. Oner: None. M. Praet: None. M. Morse: None. S. Altintas: None. S. Koljenovic: None. N. Canturk: None. P. van Dam: None. G.R. Devi: None. Background: Neoadjuvant endocrine therapy (NET) is widely used for ER⁺/HER2-negative breast cancer as it reduces tumor burden and modulates the tumor microenvironment. However, its effects on key immune cell populations remain unclear. Methods: In this retrospective study, pre- and post-treatment samples from 44 ER⁺/HER2-negative breast cancer patients treated at Antwerp University Hospital and Duke University(2006-2018) were analyzed by immunohistochemistry. We quantified stromal tumor-infiltrating lymphocytes (sTILs) along with markers for T cells, regulatory T cells (including the FOXP3/CD4 ratio), and macrophages (CD68), as well as immune checkpoint markers (PD-L1, PD-1, CTLA-4, TIM-3, and LAG-3). Treatment response was evaluated by changes in tumor size, Ki-67, and the Residual Cancer Burden (RCB) index. Results: NET significantly reduced tumor size (median decrease: 4.5 mm; p < 0.001) and Ki-67 levels (median reduction: 9%; p < 0.001) while sTIL levels remained stable (median: 5%; p = 0.08). CD4⁺ T-cell infiltration increased significantly post-treatment (p = 0.03). CD8⁺ T cells exhibited a heterogeneous response with a significant negative correlation between CD8⁺ counts and tumor size (Spearman r = -0.45, p = 0.003) indicating that higher CD8⁺ infiltration is associated with smaller tumors. FOXP3⁺ T-cell counts decreased from a median of 22.5% to 15% (p = 0.14) and the FOXP3/CD4 ratio declined significantly from a median of 2.307 to 1 (p = 0.04) suggesting a shift toward a less immunosuppressive tumor microenvironment. Higher pre-treatment CD68 expression correlated significantly with improved tumor NET response (p = 0.03). Immune checkpoint analysis revealed non-significant trends toward increased expression of PD-L1 (9% to 25%; p = 0.088), PD-1 (18% to 34%; p = 0.16), and LAG-3 (43% to 66%; p = 0.22) while CTLA-4 (23% to 27%; p = 0.69) and TIM-3 (64% to 52%; p = 0.53) remained largely unchanged. Conclusion: NET elicits profound alterations in the tumor microenvironment of ER⁺/HER2-negative breast cancer. The therapy reduces tumor proliferation and reconfigures immune cell dynamics, as evidenced by increased CD4⁺ T-cell infiltration, a beneficial reduction in the FOXP3/CD4 ratio, and a heterogeneous response of CD8⁺ T cells where higher CD8⁺ levels are linked to smaller tumors. In addition, elevated pre-treatment macrophage levels are predictive of improved tumor NET response. These results underscore the potential of integrating immune parameters as predictive biomarkers for NET efficacy and support further exploration of combined treatment strategies, including the use of immune checkpoint inhibitors, to optimize patient outcomes.Department of Defense Breast Cancer Breakthrough level 2 Award W81XWH2010153 (G.R.D.) Presentation: Monday, July 14, 2025

Background: Inflammatory breast cancer (IBC) is a rare but highly aggressive subtype of breast cancer. The tumor microenvironment (TME) plays an important role in its clinical phenotype, evolution and outcome. Cytotoxic CD8+ T cells (Tcyt) are linked to a better prognosis, while CD163+ tumor-associated macrophages (TAMs) seem to influence the rapid tumor progression of IBC and correlate with an unfavorable prognosis. Usually, immune cells in the TME are evaluated on the whole slide, but the methods used may differ significantly. Moreover, not only the quantity, but also the spatial localization of immune cells seems to be important. Therefore, we analyzed the density of CD8+ and CD163+ cells after heatmaps were generated based on their localization. Focusing only on the densities of hot spots and cold spots could increase standardization and replace a time-consuming evaluation of the whole slide. Our aim is to demonstrate the feasibility of this approach and evaluate the correlation of this heatmap with clinicopathological characteristics and outcome parameters. Method: In this retrospective cohort study, data were collected as part of an international collaboration since 1996. We included 142 patients with IBC who received their initial diagnosis and started treatment between June 1996 and December 2016. First, consecutive FFPE slides were stained with validated antibodies and digitized for further evaluation and quality control. Next, an image analysis algorithm in Visiopharm marked DAB+ immune cells and localized them spatially with XY coordinates. We then generated heatmaps showing hot and cold spots as areas of highest and lowest density (using the 90th and 10th percentile) and finally calculated the overlap of the CD8 and CD163 hot spots. Results: Most tumors were hormone receptor (HR) positive (60%, 85/142). The median sTIL score of this cohort was 12.5% (95% CI [1-80]) and 39% of the samples were PDL1 positive. Evaluation of the density in only the hot spots of both CD8 and CD163 showed a very strong correlation compared to the density of the whole slide (ρ= 0.91 and ρ= 0.92 respectively, P< .001). The presence of CD163+ TAMs correlated with more aggressive disease characteristics, both when evaluated with hot spots (HS) and on the whole slide (WS): nodal involvement (HS: P= .005 & WS: P= .04), higher tumor grade (HS: P< .001 & WS: P= .005), HR negativity (HS: P= .02 & WS: P= .04) and PDL1 positivity (HS: P< .001 & WS: P< .001). In contrast, CD8+ Tcyt correlated only with PDL1 positivity and more sTIL, and there was also no difference in evaluation of the WS or HS. Both pathological complete response (pCR) and disease-free survival (DFS) were evaluated in 108 patients that presented without metastatic disease at diagnosis and received neoadjuvant chemotherapy (NACT). Patients who achieved pCR after NACT had more CD8+ and CD163+ cells, regardless of whether evaluation was done using the WS (CD8: P= .02 / CD163: P= .006) or only the HS (CD8: P= .03 / CD163: P= .01). Finally, DFS was evaluated after dichotomizing the immune parameters. CD163+ TAMs were associated with shorter DFS (HR: 0.58, 95% CI [0.34-0.99], P= .04). This was borderline not significant in the HS analysis (HR: 0.60, 95% CI [0.34-1.05], P= .07). Although the presence of CD8+ Tcyt was not associated with longer DFS, an increasing overlap of the HS of CD8 with the HS of CD163 was associated with shorter DFS (HR: 0.59, 95% CI [0.33-1.03], P= .06), indicating the importance of immune cell interactions. Conclusion: Using heatmaps, we evaluated the density of CD8 and CD163 in both hot spots and the whole slide. There was a strong correlation between the two evaluations, proving the feasibility and reproducibility of this approach. Moreover, the presence of CD163+ TAMs was associated with more aggressive disease characteristics and worse prognosis in IBC, both when using hot spot or whole slide evaluation. Citation Format: Christophe Van Berckelaer, Isaac De Decker, Chloe Vermeulen, Peter Van Dam, Steven Van Laere, François Bertucci, Mark Kockx, Gayathri R Devi, Christophe Van Berckelaer. THE USE OF IMMUNE CELL HEATMAPS IN UNRAVELLING THE TUMOR MICROENVIRONMENT IN INFLAMMATORY BREAST CANCER [abstract]. In: Proceedings of the San Antonio Breast Cancer Symposium 2024; 2024 Dec 10-13; San Antonio, TX. Philadelphia (PA): AACR; Clin Cancer Res 2025;31(12 Suppl):Abstract nr P1-04-26.

Background: Inflammatory breast cancer (IBC), a rare and NIH-designated cancer health disparity, is a highly aggressive form of breast cancer accounting for 7-10% of all breast cancer deaths. Furthermore, the unique presentation of IBC, lacking a solid mass and characterized by diffuse tumor cell spread, often leads to delays in diagnosis and treatment. Reproductive risk factors and higher incidence rates among younger women and those from minoritized and marginalized populations, underscore the crucial role of primary care providers (PCP)—including physicians, physician assistants, and nurse practitioners. Social drivers of health (SDoH) further limit healthcare access for these communities. Recognizing that primary care practices are the initial contacts for navigating cancer diagnosis and treatment, we postulated that understanding barriers experienced by PCP and members of the public will foster engagement with large academic medical centers and breast clinics for improved diagnosis and management of IBC. In this study, we describe the development of a platform (ARCHER-C), designed to assess PCP and public knowledge, attitudes and health seeking practices related to IBC in the rural setting, as well as rare cancers more broadly. Methods: Two surveys were developed with guidance from a Community Advisory Board, comprising members from academic institutions, urban and rural practices, topical experts, and patient advocates. PCP were recruited from FQHC and community practice settings within North Carolina. Mixed methods were used to analyze cognitive interview surveys administered to a convenience sample of participants recruited at-large from rural communities via email, social media; and direct recruitment from an urban practice serving primarily low-income, medically complex patients along with another clinic serving distinct rural communities. All responses were summarized using descriptive statistics. Results: Data from public (n=34) volunteers self-reporting as Black (45%), White (30%), American Indian/Alaskan Native (24%), and residing in households earning under $50,000/yr (40%) along with PCP (n=30) were analyzed. Salient measures of IBC diagnosis, health disparity factors, referral and care coordination practices, COVID-19 impact, and continuing medical education (CME) were identified barriers at the PCP level. Furthermore, seventy-four percent of public participants had not heard of IBC; 38% did not identify being overweight as a risk factor; 38% were not aware of racial disparities; 50% recognized redness or thickening of the skin as symptoms; and even fewer recognized the characteristic “pitted” appearance of the skin. Participants noted worry about what the doctor may find, difficulty or fear talking to a physician, lack of insurance, transportation barriers, and religious reasons as contributing to delays seeking health care. Most importantly, 97% of the participants expressed their enthusiasm in sharing information about IBC and rare cancers with others, specifically family members and friends. In addition, majority expressed that adding visual aids and providing educational materials about IBC at the time of survey would be helpful. Conclusions: This community-engaged model highlights the need for bidirectional provider- and public-facing research and education to reduce the burden of IBC and associated disparities. Strengthening the connection between primary care and specialized centers is essential for better diagnosis and management of IBC and other rare cancers, ultimately reducing health disparities. Funding in part by Duke Cancer Institute Community Outreach, Engagement, Equity Seed Grant (GRD, ANT) as part P30 CA014236, Duke MERITS Education Grant & Surgery Funds (GRD). Anh Tran and Gayathri Devi; Co-Senior Authors. Citation Format: Beau Blass, Hannah Woriax, Rashmi Saincher, Larry Greenblatt, Anh N. Tran, Gayathri R. Devi. Beau Blass, Hannah Woriax, Rashmi Saincher, Larry Greenblatt, Anh N. Tran, Gayathri R. Devi [abstract]. In: Proceedings of the San Antonio Breast Cancer Symposium 2024; 2024 Dec 10-13; San Antonio, TX. Philadelphia (PA): AACR; Clin Cancer Res 2025;31(12 Suppl):Abstract nr P3-03-01.

Therapeutic resistance presents a significant hurdle in combating inflammatory breast cancer (IBC), adding to the complexity of its management. To investigate these mechanisms, we conducted a comprehensive analysis using transcriptomic and proteomic profiling in a preclinical model alone with correlates of treatment response in IBC patients. This included SUM149 cell lines derived from treatment-naïve patients, along with acquired drug resistance (rSUM149) and others in a state of resistance reversal (rrSUM149), aiming to uncover drug resistance networks. We identified specific ribosomal proteins associated with acquiring resistance. These correlated with elevated levels of molecular markers such as pERK, CDK1, XIAP, and SOD2. While resistance reversal in rrSUM149 cells largely normalized the expression profile, VIPER analysis revealed persistent alterations in ribosomal process-related proteins (AGO2, Exportin 1, RPL5), suggesting their continued involvement in drug resistance. Moreover, genes linked to ribosomal processes were significantly enriched (P < 0.001) among overexpressed genes in IBC patients (n = 87) who exhibited a pathological complete response (pCR) to neoadjuvant chemotherapy. Given the common hyperactivation of MAPK in IBC tumors, including rSUM149, we evaluated Merestinib, a multikinase inhibitor in clinical trials. It effectively targeted pERK and peIF4E pathways, suppressed downstream targets, induced cell death in drug-resistant rSUM149 cells, and showed synergistic effects with another tyrosine kinase inhibitor (Lapatinib) in parental cells. This underscores its significant impact on protein synthesis signaling, crucial for combating translational dependence in cancer cells. In summary, our study elucidates adaptive changes in IBC cells in response to therapy and treatment pauses, guiding precision medicine approaches for this challenging cancer type.

Abstract Background: Inflammatory breast cancer (IBC) is distinguished by the presence of tumor emboli, clusters of tumor cells, found within breast parenchyma and dermal lymphatic vessels. These emboli serve as a crucial clinicopathological hallmark, believed to underlie the characteristic diffuse growth pattern and aggressive progression observed in IBC patients, with nearly 30% of patients presenting with metastatic disease at diagnosis. However, the rarity of this cancer and the absence of a solid tumor mass pose challenges in obtaining biospecimens, particularly those with evidence of tumor emboli. To address this pressing need for preclinical models that replicate this distinctive tumor growth pattern, we describe a tumor emboli culture platform that facilitates live imaging, enables compound screening by assessing various phenotypes of the emboli, and permits evaluation of emboli in an in vivo murine model. Methods: The tumor emboli culture system was evaluated for physiological measurements corresponding to dermal lymphatic vessels and subsequently employed to generate tumor emboli from diverse patient-derived cell lines and PDX. Confocal microscopy was employed to characterize cellular information across multiple slices of the tumor emboli. This provided a comprehensive, multi-parametric analysis of single-cell and spheroid phenotypes, including measurements of number, size, shape, and viability of the organoids. Furthermore, we developed imaging algorithms to evaluate phenotypic changes in tumor emboli in response to anti-cancer drug treatment. We also optimized a technique that allows for in vivo tumor emboli implantation including in a transgenic mice with red fluorescent lymphatic vasculature that were surgically implanted with a dorsal skin window chamber. This allowed for intravital optical imaging of the local tumor microenvironment and adjacent skin. Results: Our culture platform enables live imaging of the tumor emboli generated over a span of 7-10 days, maintaining key parameters including kinematic and dynamic viscosity, density along with sheer wall stress within the reported physiological range for lymphatics. Through higher resolution confocal image acquisition and multi-parametric analysis, we conducted particle counting and classification, providing statistically significant insights into the integrity of tumor emboli over time. Additionally, the developed algorithms allowed for quantitative analysis of images from tumor emboli cultures subjected to anti-cancer drugs, revealing disparities in drug efficacy on tumor emboli integrity and dispersion including inter-cluster space within the emboli. Intravital optical imaging, coupled with quantitative analysis of dispersion characteristics of the tumor emboli implanted in the murine model over 5-6 days, demonstrated a diffuse spread reminiscent of observations in IBC patients. Furthermore, immunohistochemistry staining showed marked Ki67, CD45, F4-80 revealing a proliferative tumor microenvironment with high levels of macrophage infiltration. Conclusions: As of now specific targeted therapies for IBC patients remain elusive. Our culture platform for tumor emboli, when combined with advanced imaging techniques both in vitro and in vivo and traditional approaches like immunohistochemistry and genomic evaluation, forms a powerful toolkit for assessing the phenotype and dispersion characteristics of this unique pathological feature in IBC. This preclinical model offers crucial insights, potentially paving the way for precision medicine strategies in the treatment of this understudied cancer. Funding in part from American Cancer Society Mission Boost MBG-20-141-01-MBG grant (GRD), Department of Defense W81XWH-20-1-0153 (GRD), Duke Consortium for Inflammatory Breast Cancer Education (AB, CW, RE). Citation Format: Dorababu Sannareddy, Theresa Charity, Alexandra Bennion, Caroline Way, Edwin Yu, Ralph Erdmann, Gregory M Palmer, Gayathri R Devi. A Tumor Emboli Platform for Morphometric Analysis, Drug screening and for Intravital Optical Imaging in a Murine Model to Simulate Clinicopathological Features of Inflammatory Breast Cancer [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO5-27-02.

Modification of catalytic expression of enzymes and regulating their in vivo activity are the goals of novel treatment strategies. A green synthetic nanostructured silver with potent trypsin inhibitory properties has not yet been developed, despite the fact that silver nanoparticles possess unique properties that allow them to efficiently block enzymes. The present study demonstrates for the first time a facile, safe, economic, and eco-friendly synthetic route for silver nanoparticles using an aqueous extract of Holigarna arnottiana bark engineered to interact with trypsin and hinder its activity effectively. The studies carried out to examine the interaction between these biofabricated AgNPs (HaAgNPs) and trypsin by UV-visible spectrophotometry and FTIR spectroscopy suggest that the formation of trypsin-HaAgNP complex is responsible for diminishing the catalytic efficiency of trypsin. In vivo studies on Aedes aegypti larval serum support these instrumental results of HaAgNP-induced trypsin inhibition and proves its application as a biopesticide. It is noteworthy that the bioengineered HaAgNPs were also found to have good inhibition potential against pepsin and urease as well. A variety of methods have been employed to characterize the synthesized biocompatible HaAgNPs and it possesses a characteristic absorption maximum of 420 nm. Their shelf life of above 7 years is noticeable, since none of the reported green synthesized AgNPs possess a shelf life of more than 1 year. Altogether, this work demonstrates that biofabricated HaAgNPs are multifunctional and cost-resilient biological tools that can be used as enzyme regulators possessing antioxidant, antimicrobial, and insecticidal features.

Abstract Background: The diagnosis of inflammatory breast cancer (IBC) is life-changing as it is a rare, highly aggressive breast cancer variant designated by NIH as a cancer health disparity. Given the unique presentation of diffuse tumor cell spread in the absence of a solid mass, which is not easily detected by self-breast exams or mammograms, IBC patients often face delays in diagnosis and treatment. Therefore, an individualized and a multifaceted approach that addresses the unique needs of the patient with input from both providers and the community is an unmet need. To address this, we are investigating the current knowledge and perspectives of providers and public related to rare cancers like IBC in primary care practices. Methods: Researchers and advanced practice providers collaboratively drafted and piloted the initial surveys with guidance from a Community Advisory Board, comprising of members from academic institutions, primary care practices in urban and rural setting, topical experts, and patient advocates, in order to assess primary care providers (PCPs) and general public knowledge, attitudes and health seeking practices related to rare cancer like IBC. Results: The surveys are currently being administered statewide specific to PCPs or public via cognitive interviews and/or using an online survey tool within North Carolina as a model of a large state (100 counties) with a range of population densities, race, ethnicities, and environmental burdens. Challenges elaborated at the PCP level include communicating rare cancer information, gaps in confidence in diagnosing IBC, timely follow-up with patients, and accessing specialty care and an unmet need for developing PCP educational modules to improve guideline-concordant care. Public data analyzed to date is from a Duke Outpatient Clinic, an urban ambulatory care teaching practice serving primarily low-income, medically complex patients in Durham (n=14) included 69% Black, 93% female, with 68% in households earning under $50,000 annually). 71% had not heard of IBC; 64% did not identify high BMI as a risk factor; 36% did not know there is differential race-related incidence; 74% did not recognize IBC-specific symptoms. 50% noted lack of insurance, transportation, religious reasons, or difficulty or fear of talking to a physician contributed in general delay in seeking health care. Most importantly, 93% expressed feeling comfortable sharing information about IBC with others. Majority expressed that adding visual aids and providing educational materials about IBC at the time of survey would be helpful. Conclusions: The results highlight the role of PCPs, including physicians, physician assistants, and nurse practitioners, who are often the first point of contact when patients begin to notice signs or symptoms. The general public and the patients with common breast cancer subtypes can provide valuable insight into their own health care experiences, what works for them and what barriers they face in their communities, which can be used to compare with experience of patients with IBC and other rare cancers. Overall, the lessons from this study inform the development of effective tools to educate PCPs and patients for this understudied cancer that contributes to disparities in clinical and survival outcomes in minoritized and marginalized populations. Support in part from the Duke Cancer Institute pilot as part of the P30 Cancer Center Support Grant (P30CA014236) and Duke School of Medicine Behavioral Health and Survey Research Core Facility Award (GRD, ANT); Duke Advanced Practice Provider Leadership Institute scholarship (VK, GRD), Department of Surgery (GRD); Duke Ahead Education grant; NIH/NCI RO1CA264529-01 (GRD); ACS Mission Boost grant MBG-20-141-01-MBG (GRD), Nakayama Public Service Scholar Award (AB). Citation Format: Gayathri R Devi, Lawrence Greenblatt, Rashmi Saincher, Valerie Keck, Beau Blass, Sarah Weaver, Alexandra Bennion, Anh Tran. Rare to Care: Assessing Awareness and Health Disparity Factors Related to Inflammatory Breast Cancer in the Primary Care Setting [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO5-10-04.

Photodynamic therapy (PDT) stands out as a noteworthy development as an alternative targeted treatment against skin ailments. While PDT has advanced significantly, research into photo-activatable “Green drugs” derived from plants which are less toxic than the synthetic drugs has not kept pace. This study investigates the potential of Fagopyrin F Containing Fraction (FCF) derived from Fagopyrum tataricum in mediating PDT against Staphylococcus aureus and skin cancer cells (A431). FCF was isolated from the plant extract using thin-layer chromatography, followed by identification of the compound through high-performance liquid chromatography and high-resolution liquid chromatography–mass spectrometry. FCF was tested to determine its antibacterial and anticancer efficacy. Results revealed that FCF-mediated PDT exhibited potent action against S. aureus, significantly reducing bacterial viability (MIC 19.5 μg/100 μL). Moreover, FCF-mediated PDT showed good efficacy against A431 cells, resulting in a notable reduction in cell viability (IC50 29.08 μg/mL). Given the known association between S. aureus and squamous cell carcinoma (SCC), FCF shows the potential to effectively target and eradicate both SCC and the related S. aureus present within the lesions. In silico study reveals that Fagopyrin F effectively binds with the epidermal growth factor (EGFR), one among the highly expressed proteins in the A431 cells, with a binding energy of − 9.6 kcal/mol. The affinity of Fagopyrin F for EGFR on A431 cancer cells along with its cytotoxicity against skin cancer cells while safeguarding the normal cells (L929) plays a major part in the way it targets cancer cells. However, its safety, efficacy, and long-term advantages in treating skin conditions require more investigation, including in vivo investigations and clinical trials.

Novel formulations of silver nanoparticles remain exciting if it is applicable for cosmetic purposes. This study proposes a value-added brand-new nanomaterial for improving skin complexion by inhibiting melanin development. This work aims to develop cost effective, efficient, natural silver nanoparticles phytomediated by aqueous extract of leaf sheath scales of Cocos nucifera (Cn-AgNPs) having potential as tyrosinase inhibitors hindering melanin synthesis. The formation of Cn-AgNPs was assessed spectrophotometrically and confirmed by the sharp SPR spectrum at 425 nm. The chemical composition profiling was characterized by X-ray diffraction (XRD) and Fourier Transform Infrared (FTIR) spectroscopy. The morphology was confirmed by Field Emission Scanning Electron Microscopy (FESEM) and the thermal stability was assessed by Thermogravimetric analysis (TGA). Pharmacological application studies supported the materialization of Cn-AgNPs with significant antityrosinase potential and considerably improved antibacterial and antioxidant properties. Cn-AgNPs showed potential antibacterial effects against gram-positive and negative strains, including prominent infectious agents of the skin. Antioxidant capacity was confirmed with an IC50 of 57.8 μg/mL by DPPH radical scavenging assay. Furthermore, in vitro melanin content determination was performed using SK-MEL cells. Cell line studies proved that Cn-AgNPs decrease the melanin content of cells. The IC50 value obtained was 84.82 μg/mL. Hence Cn-AgNPs is proposed to be acting as a whitening agent through lessening cellular melanin content and as a significant inhibitor of tyrosinase activity. The antioxidant properties and antibacterial effects can contribute to skin rejuvenation and can prevent skin infections as well. This evidence proposes the development of a new nanostructured pharmaceutical and cosmetic formulation from Cocos nucifera leaf sheath scales.

Individual environmental contaminants have been associated with breast cancer; however, evaluations of multiple exposures simultaneously are limited. Herein, we evaluated associations between breast cancer summary stages and the Environmental Quality Index (EQI), which includes a range of environmental factors across five domains. The EQI (2000–2005) was linked to county-level age-standardized incidence rates (SIRs) obtained from the North Carolina Central Cancer Registry (2010–2014). Incidence rates and SIRs of total, in situ, localized, regional, and distant breast cancers were evaluated stratified by rural–urban status. In counties with poor environmental quality compared to those with good environmental quality, total breast cancer incidence was higher by 10.82 cases per 100,000 persons (95% CI 2.04, 19.60, p = 0.02). This association was most pronounced for localized breast cancer (β = 5.59, 95% CI 0.59, 10.58, p = 0.03). Higher incidence of early-stage disease (carcinoma in situ β = 5.25, 95% CI 2.34, 8.16, p = 0.00 and localized breast cancer β = 6.98, 95% CI 2.24, 11.73, p = 0.00) and total breast cancer (β = 11.44, 95% CI 3.01, 19.87, p = 0.01) occurred in counties with poor land quality, especially urban counties. Our analyses indicate significant associations between environmental quality and breast cancer incidence, which differ by breast cancer stage and urbanicity, identifying a critical need to assess cumulative environmental exposures in the context of cancer stage.

Inflammatory breast cancer (IBC), an understudied and lethal breast cancer, is often misdiagnosed due to its unique presentation of diffuse tumor cell clusters in the skin and dermal lymphatics. Here, we describe a window chamber technique in combination with a novel transgenic mouse model that has red fluorescent lymphatics (ProxTom RFP Nu/Nu) to simulate IBC clinicopathological hallmarks. Various breast cancer cells stably transfected to express green or red fluorescent reporters were transplanted into mice bearing dorsal skinfold window chambers. Intravital fluorescence microscopy and the in vivo imaging system (IVIS) were used to serially quantify local tumor growth, motility, length density of lymph and blood vessels, and degree of tumor cell lymphatic invasion over 0–140 h. This short-term, longitudinal imaging time frame in studying transient or dynamic events of diffuse and collectively migrating tumor cells in the local environment and quantitative analysis of the tumor area, motility, and vessel characteristics can be expanded to investigate other cancer cell types exhibiting lymphovascular invasion, a key step in metastatic dissemination. It was found that these models were able to effectively track tumor cluster migration and dissemination, which is a hallmark of IBC clinically, and was recapitulated in these mouse models.