Healthy Breast Research Articles

Elemental Characterization and Radiation Parameters of Malignant and Healthy Breast Tissues

IntroductionElemental imbalances in breast cancer patients have been linked with carcinogenic processes which are responsible for the development and growth of breast malignancy. In medical applications of radiation, this elemental characterization is important in determining the radiation parameters which are fundamental in understanding the interaction of radiation with concerned tissue. This study aims to carry out an elemental characterization of malignant and healthy breast tissue and to determine their respective radiation parameters. MethodsParticle-induced X-ray emission (PIXE) procedure was used for the elemental characterization of the samples. The resulting elemental concentrations were used in calculating the photon radiation parameters with the aid of the “EpiXS” software. ResultsAmong the elements determined, Mg, Al, P, S, Cl, K, Mn, Zn, and Pb in the malignant group were found to be significantly higher than that of the healthy group at a 0.05 significance level. Although not significant, the effective atomic number (Zeff) and the electron density (ρe) of the malignant group were slightly higher than that of the healthy group and this causes the difference in the mass attenuation coefficients for photoelectric effect and pair production interactions of the two subjects. The healthy tissue which has the lower value of Zeff and ρe dominated the maximum values of EABF while the minimum values were dominated by the malignant breast tissue which has the higher Zeff and ρe. The variation in the photon radiation parameters between the two groups suggests that the two groups will behave differently when photons interact with them. ConclusionThe results obtained in this study will be helpful in radiation biological physics and thus be proven worthwhile in radiodiagnostic, radiation dosimetry, and radiation therapy of breast cancer.

Nondestructive protein sampling with electroporation facilitates profiling of spatial differential protein expression in breast tumors in vivo

Excision tissue biopsy, while central to cancer treatment and precision medicine, presents risks to the patient and does not provide a sufficiently broad and faithful representation of the heterogeneity of solid tumors. Here we introduce e-biopsy—a novel concept for molecular profiling of solid tumors using molecular sampling with electroporation. As e-biopsy provides access to the molecular composition of a solid tumor by permeabilization of the cell membrane, it facilitates tumor diagnostics without tissue resection. Furthermore, thanks to its non tissue destructive characteristics, e-biopsy enables probing the solid tumor multiple times in several distinct locations in the same procedure, thereby enabling the spatial profiling of tumor molecular heterogeneity.We demonstrate e-biopsy in vivo, using the 4T1 breast cancer model in mice to assess its performance, as well as the inferred spatial differential protein expression. In particular, we show that proteomic profiles obtained via e-biopsy in vivo distinguish the tumors from healthy breast tissue and reflect spatial tumor differential protein expression. E-biopsy provides a completely new molecular sampling modality for solid tumors molecular cartography, providing information that potentially enables more rapid and sensitive detection at lesser risk, as well as more precise personalized medicine.

Single cell atlas identifies lipid-processing and immunomodulatory endothelial cells in healthy and malignant breast

Since a detailed inventory of endothelial cell (EC) heterogeneity in breast cancer (BC) is lacking, here we perform single cell RNA-sequencing of 26,515 cells (including 8433 ECs) from 9 BC patients and compare them to published EC taxonomies from lung tumors. Angiogenic ECs are phenotypically similar, while other EC subtypes are different. Predictive interactome analysis reveals known but also previously unreported receptor-ligand interactions between ECs and immune cells, suggesting an involvement of breast EC subtypes in immune responses. We also identify a capillary EC subtype (LIPEC (Lipid Processing EC)), which expresses genes involved in lipid processing that are regulated by PPAR-γ and is more abundant in peri-tumoral breast tissue. Retrospective analysis of 4648 BC patients reveals that treatment with metformin (an indirect PPAR-γ signaling activator) provides long-lasting clinical benefit and is positively associated with LIPEC abundance. Our findings warrant further exploration of this LIPEC/PPAR-γ link for BC treatment.

Metabolomics links lifestyle variables to breast cancer in the Long Island Breast Cancer Study Project

Background/Aim: Healthy lifestyle has been associated with decreased risk of developing breast cancer. Using untargeted metabolomics profiling, we aim to identify the molecular mechanisms connecting lifestyle and breast cancer through network analysis. Methods: A total of 50 post-menopausal women with breast cancer and 50 cancer-free controls were selected from the Long Island Breast Cancer Study Project (LIBCSP). We measured untargeted plasma metabolomics using liquid chromatography- high resolution mass spectrometry (LC-HRMS). Using ‘enet’, we retained metabolites found in active molecular network (AMN) clusters for analysis. LASSO was used to explore associations between cancer status and metabolites; 14 lifestyle related variables including smoking, physical activity, alcohol consumption, meat consumption, fruit and vegetables consumption, supplemental vitamin use; and 20 covariates including various reproductive factors. LASSO was repeated to explore associations between selected lifestyle factors and metabolites. Results were displayed as a network to uncover biological pathways linking lifestyle factors to breast cancer. Results: After filtering, 1797 metabolomics peaks in the plasma samples remained. Of these, 851 "active” metabolites were retained in 197 correlation AMN clusters. Through LASSO, breast cancer status was associated with 66 "active” metabolites. Several of these metabolites were also associated with lifestyle variables including meat consumption, alcohol consumption, and supplemental β-carotene, B12 and folate use. No individual lifestyle factors were directly associated with breast cancer status through LASSO, suggesting that metabolites may act as biological intermediaries between healthy lifestyle factors and breast cancer. In particular, 13-HPODE, a metabolite linked with inflammation, was associated with breast cancer status and connected to β-carotene supplement usage through an AMN. Conclusions: We found several plasma metabolites associated with both lifestyle factors and breast cancer status. These results suggest that inflammation may play a mechanistic role linking supplement usage to breast cancer status. Keywords: breast cancer, lifestyle factors, metabolomics, network analysis

In vitro effects of combustion generated carbon dots on cellular parameters in healthy and cancerous breast cells

Carbon nanomaterials are an inventive class of materials with wide applications in state-of-the-art bioimaging and therapeutics. They allow a broad range of tunable and integrated advantages of structural flexibility, chemical and thermal stability, upright electrical conductivity, and the option of scale-up and mass production. In the context of nanomedicine, carbon nanomaterials have been used extensively to mitigate the serious side effects of conventional chemotherapy and also to enable early cancer diagnostics, given their wide range of tunable properties. A class of carbon nanomaterials, called carbon dots (CDs) are small carbon-based nanoparticles and have been a valued discovery due to their photoluminescence, low photobleaching, and high surface area to mass ratio. The process of producing these CDs had so far been a high energy demanding process involving wet chemistry for purification. A one-step tunable production of luminescent CDs from fuel rich combustion reactors was recently presented by our group. In this paper, we explore the effects of these yellow luminescent combustion-generated CDs in MCF7 adenocarcinoma and MCF10a normal breast epithelial cells. We observed that these CDs, also at nontoxic doses, can affect basic cellular functions, such as cell cycle and proliferation; induce substantial changes on the physical parameters of the plasma membrane; and change the overall appearance of a cell in terms of morphology.

Identification of the phenomenon of anastasis in breast cancer cells

Amaç: Çalışmada, dünya genelinde en yaygın kanser tiplerinden biri olan meme kanserinde, apoptoz ile ilişkili genlerin anastaz sürecindeki ifadesel değişimlerinin ve potansiyel rollerinin tanımlanması amaçlanmıştır.
 Gereç ve Yöntem: Farklı tipteki meme kanseri hücre hatları (MCF7 ve MDA-MB-231), meme kanseri kök hücreleri ve sağlıklı meme hücre hattı (MCF10A) kullanıldı. Apoptotik ve anastatik hücre yüzdeleri, akış sitometrisi aracılığıyla Annexin V testi ile belirlendi. Apoptotik hücrelere göre anastatik hücrelerin gen ekspresyon değişimleri qRT-PCR ve 2-ΔΔCt yöntemi ile belirlendi. Anlamlı değişim gösteren genlerin görev aldıkları yolak ve biyolojik süreçler STRING v11.5 veri tabanı kullanılarak belirlendi.
 Bulgular: Tüm hücre gruplarında etanol uygulaması sonucu kontrole göre apoptoz yüzdesinin arttığı ve apoptozu uyaran etmenin uzaklaştırılmasıyla apoptotik hücre yüzdesinin azaldığı belirlendi. Kontrol, apoptoz ve anastaz grupları arasında apoptotik hücre yüzdesindeki değişim en fazla MCF7 hücrelerinde belirlendi. Uyumlu şekilde bu hücre hattında en fazla sayıda gende ifade değişimi belirlendi. CASP7 ve APAF1 genleri tüm hücre hatlarında ekspresyon azalışı sergiledi. Tüm hücre gruplarında anastazın sistein tipi endopeptidaz aktivitesini (GO_ID: 0043027) ve ilaç direnci ilişkili yolakları (KEGG_ID: hsa01524) ortak şekilde etkilediği belirlendi.
 Sonuç: Hücrelerde anastaz fenomeninin apoptoz düzenleyici mekanizmalar ile etkileşiminin tanımlanması, hem sağlıklı hücrelerin onkojenik dönüşümünün hem de kanserde ilaç direnci mekanizmalarının aydınlatılabilmesi açısından önem taşımaktadır.

Operating on Anxiety: Negative Affect toward Breast Cancer and Choosing Contralateral Prophylactic Mastectomy.

Rates of contralateral prophylactic mastectomy (CPM)-removal of the healthy breast following breast cancer diagnosis-have increased, particularly among women for whom CPM provides no survival benefit. Affective (i.e., emotional) decision making is often blamed for this increase. We studied whether greater negative breast cancer affect could motivate uptake of CPM through increased cancer risk perceptions and biased treatment evaluations. We randomly assigned healthy women with average breast-cancer risk (N = 1030; Mage = 44.14, SD = 9.23 y) to 1 of 3 affect conditions (negative v. neutral v. positive narrative manipulation) in a hypothetical online experiment in which they were asked to imagine being diagnosed with cancer in one breast. We assessed 1) treatment choice, 2) affect toward CPM, and 3) perceived risk of future breast cancer in each breast (cancer affected and healthy) following lumpectomy, single mastectomy, and CPM. The manipulation caused women in the negative and neutral narrative conditions (26.9% and 26.4%, respectively) to choose CPM more compared with the positive narrative condition (19.1%). Across conditions, women's CPM affect did not differ. However, exploratory analyses addressing a possible association of affect toward cancer-related targets suggested that women in the negative narrative condition may have felt more positively toward CPM than women in the positive narrative condition. The manipulation did not have significant effects on breast cancer risk perceptions. The manipulation of affect had a small effect size, possibly due to the hypothetical nature of this study and/or strong a priori knowledge and attitudes about breast cancer and its treatment options. Increased negative affect toward breast cancer increased choice of CPM over other surgical options and might have motivated more positive affective evaluations of CPM. This study used narratives to elicit different levels of negative integral affect toward breast cancer to investigate the effects of affect on breast cancer treatment choices.Increased negative affect toward breast cancer increased the choice of double mastectomy over lumpectomy and single mastectomy to treat a hypothetical, early-stage cancer.The narrative manipulation of negative affect toward breast cancer did not change the perceived risks of future cancer following any of the surgical interventions.Negative affect toward breast cancer may have biased affective evaluations of double mastectomy.

Determination of adenosine and its modifications in urine and plasma from breast cancer patients by hydrophilic interaction liquid chromatography-tandem mass spectrometry

RNA modifications have been revealed to be essential in many biological activities, and their disorders are associated with various human diseases, including cancers. 2′-O-methyladenosine (Am), N1-methyladenosine (m1A), N6-methyladenosine (m6A), N6,2′-O-dimethyladenosine (m6Am) and N6,N6-dimethyladenosine (m62A) are important adenosine (A) modifications. The noninvasive collection of urine samples and the diverse contents of metabolites in plasma make them favored biofluids for biomarkers discovery. In this work, we established a hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) method to quantify these six nucleosides in urine and plasma of healthy controls and breast cancer (BC) patients. The limit of detection (LOD) for A, Am, m1A, m6A, m6Am, and m62A were 0.0025, 0.01, 0.05, 0.005, 0.005, and 0.005 nM. The results showed that the concentrations of Am, m6A, and m6Am were increased, whereas m1A was decreased in the urine of BC patients compared with the healthy controls. We also found that the level ratios of m1A/A, m6A/A, and m6Am/A were all reduced in plasma from BC patients, compared with healthy controls. Interestingly, these ratios of methylated adenosine nucleosides to adenosine in plasma could better discriminate BC patients from healthy controls, compared to the levels of these nucleosides. The present study not only suggests these modified adenosines can act as noninvasive biomarkers of BC but also will contribute to investigating the impacts of RNA methylation on the occurrence and development of BC.

Flap Monitoring Using Interstitial Fluid Glucose Measurements.

Background: Various flap monitoring techniques have been used in the early detection of anastomotic thrombus; however, the use of automatic and continuous monitoring methods is presently uncommon. The purpose of this study was to investigate trends in interstitial fluid glucose concentration (IFG) in flap monitoring by measuring IFG automatically and continuously. Methods: Nine patients underwent unilateral breast reconstruction using a transverse rectus abdominis myocutaneous flap with vascular anastomosis. Two IFG measuring devices were attached to each patient. One device was attached to the flap (flap IFG) and the other to the healthy breast (control IFG). In each case, flap IFG, control IFG, and IFG ratio (flap IFG/control IFG) were recorded in the initial 72 h post-surgery (first half) and also in the subsequent 72 h (second half). In all of the cases, the mean values recorded in the first half and those in the second half were compared. Results: All flaps survived. The flap IFG didn't fall below 40 mg/dL in the first half. The minimum flap IFG and IFG ratio were 42 mg/dL and 0.55 in the first half. The flap IFG was significantly higher in the first half than in the second half in all cases, and the IFG ratio was similar in 8 cases. Furthermore, mean flap IFG and mean IFG ratio in all cases in the first half were significantly higher than in the second half. Conclusions: IFG measurements may complement conventional flap monitoring, particularly in the early postoperative period.

Single-Molecule Counting of FTO in Human Breast Tissues Based on a Rolling Circle Transcription Amplification-Driven Clustered Regularly Interspaced Short Palindromic Repeat─Cas12a.

N6-methyladenosine modification as an mRNA modification in mammalian cells is dynamically reversible, regulated by RNA demethylase [e.g., fat mass and obesity-associated protein (FTO)]. The abnormal expression of FTO is closely related to numerous diseases (e.g., various cancers and obesity). Herein, we demonstrate the single-molecule counting of FTO in human cancer cells and breast tissues based on a T7 RNA polymerase-mediated rolling circle transcription (RCT) amplification-driven clustered regularly interspaced short palindromic repeat (CRISPR)─Cas12a. When FTO is present, it demethylates the DNA substrate, initiating the DpnII-mediated cleavage reaction. After magnetic separation, the cleaved DNA fragments trigger the T7 RNA polymerase-mediated RCT amplification, activating CRISPR-/Cas12a-mediated cleavage of signal probes and releasing abundant FAM molecules that are simply counted via single-molecule detection. In this assay, only target FTO can generate CRISPR RNAs, efficiently improving detection specificity. Moreover, the integration of single-molecule detection with magnetic separation achieves zero background and effectively enhances detection sensitivity. This method can specifically and sensitively monitor FTO activity with a limit of detection of 1.20 × 10-13 M, and it may measure FTO at the single-cell level. Furthermore, it may accurately discriminate the FTO expression level in breast tissues between healthy persons and breast cancer patients and screen the FTO inhibitors as well, with great potential in clinical diagnosis and drug discovery.

A New Technique for Breast Pocket Adjustment: Argon Beam Thermal Capsulorrhaphy.

Summary:Implant malposition remains one of the main complications of aesthetic breast augmentation and alloplastic breast reconstruction with expanders and implants. Many capsulorrhaphy techniques have been described to adjust the breast pocket and correct the malposition. In this study, we tested the efficacy of the argon beam coagulator (ABC) for lateral capsulorrhaphy on breast reconstruction patients at the time of expander replacement with a permanent implant. We also experimentally compared the effects of the ABC and the standard electrocautery on fragments of healthy breast capsule. We noted a 69.5% capsule shrinkage with the ABC versus 46.8% with the standard electrocautery. We concluded that breast capsulorrhaphy using the ABC is a safe and efficient technique for the correction of breast implant malposition in both reconstructive and aesthetic breast surgery.

Lapatinib loaded exosomes as a drug delivery system in breast cancer

Nanocarriers and nanoparticles in the drug delivery systems increase the drug's effectiveness due to their permeability and retention properties to tumor tissues. Exosomes are small extracellular natural vesicles released from the plasma membrane and they promise drug delivery vehicles to be used in anti-cancer therapy. The prominent features of exosomes as nanocarriers are biocompatibility, their natural form, high stability, and low toxicity. These features also distinguish the exosome from synthetic nanoparticles. Exosomes were isolated from healthy epithelial breast cells (MCF10 A) using an aqueous two-phase system (ATPS). Isolated exosomes were characterized with NTA (Nanoparticle Tracking Analysis), Zeta sizer, AFM (Atomic Force Microscopy), and flow cytometry. Lapatinib chemotherapeutic drug was loaded into isolated exosomes with the electroporation method. The loaded drug was characterized and measured using high-performance liquid chromatography (HPLC). The cytotoxicity of free lapatinib (Lap), lapatinib encapsulated exosome (LapExo), and empty exosome (Exo) were determined in HER2 positive breast cancer SKBR 3 and healthy breast MCF10 A cells. Lapatinib-loaded exosomes significantly reduced cellular proliferation of SKBR 3 cells at lower doses compared to free Lap treatment. Apoptosis analysis showed that LapExo treatment led to a high apoptotic rate in comparison to free Lap treatment on SKBR 3 cells. Lapatinib targeting mechanisms through the downstream components of HER2 and EGFR signaling pathway were evaluated post Lap and LapExo treatment on breast cancer SKBR-3 and healthy MCF10 A cells. LapExo treatment showed similar effect on mRNA expression and protein levels as Lap treatment. This indicates that exosomal loading and delivery of chemotherapeutic drugs did not interfere the acting mechanism of drug. In the present study, we successfully loaded lapatinib chemotherapeutic drugs in exosomes that derived from non-cancerous epithelial breast cells. The lapatinib-loaded exosomes allowed us to use the lower dose concentration of the drug for breast cancer treatment. They increased the effectiveness of drug compared to lapatinib alone. Overall, this study revealed that using of exosome natural vesicles in drug delivery approaches enable cancer therapy to be more effective and could enhance the therapeutic index of chemotherapeutic drugs.

Target selectivity of cholesterol-phosphatidylcholine liposome loaded with phthalocyanine for breast cancer diagnosis and treatment by photodynamic therapy

This study investigated the ability of cholesterol-phosphatidylcholine liposomes loaded with chloride aluminum phthalocyanine (CL-AlClPc) to discriminate between healthy (MCF-10A) and neoplastic (MCF-7 and MDA-MB-231) breast cells for breast cancer diagnosis and treatment by photodynamic therapy (PDT) using a new drug delivery system consisting of CL-AlClPc. When PDT treatment was applied at an energy fluence of 700 mJ/cm², CL-AlClPc was more cytotoxic to neoplastic cells than to healthy breast cells because CL-AlClPc was better internalized by the tumor cells. An even higher fluorescence signal is expected for neoplastic cells during clinical treatment than for healthy cells, which will be useful for precise and targeted tumor cell detection. CL-AlClPc also facilitated better drug distribution and targeting of essential organelles inside the cells. This selectivity is critical for future in vivo diagnosis and treatment; it prevents side effects because it prioritizes tumor cells and tissues instead of healthy ones. The CL-AlClPc system designed herein had a small size (150 nm), low zeta potential (-6 mV), low polydispersity (0.16), high encapsulation rate efficiency (82.83%), and high shelf stability (12 months).

Profiling of mature-stage human breast milk cells identifies six unique lactocyte subpopulations.

Breast milk is chock-full of nutrients, immunological factors, and cells that aid infant development. Maternal cells are the least studied breast milk component, and their unique properties are difficult to identify using traditional techniques. Here, we characterized the cells in mature-stage breast milk from healthy donors at the protein, gene, and transcriptome levels. Holistic analysis of flow cytometry, quantitative polymerase chain reaction, and single-cell RNA sequencing data identified the predominant cell population as epithelial with smaller populations of macrophages and T cells. Two percent of epithelial cells expressed four stem cell markers: SOX2, TRA-1-60, NANOG, and SSEA4. Furthermore, milk contained six distinct epithelial lactocyte subpopulations, including three previously unidentified subpopulations programmed toward mucosal defense and intestinal development. Pseudotime analysis delineated the differentiation pathways of epithelial progenitors. Together, these data define healthy human maternal breast milk cells and provide a basis for their application in maternal and infant medicine.

A Breast Cancer Detection System Using Metasurfaces With a Convolution Neural Network: A Feasibility Study

This work presents a breast cancer detection modality using a metasurface as the imaging medium and a microwave source. The fundamental concept is to record the electromagnetic energy, which, when transmitted through the breast, scatters differently according to the variation in the electrical properties of the breast tissue. The energy captured by the metasurface forms an impression image of the breast under test. This impression is used as a detection tool to determine the presence of a tumor inside the breast. A convolution neural network (CNN) is applied to enhance detection accuracy and provide quantitative data about tumors when present. The CNN results show the possibility of finding a tumor with a 10 mm diameter and distinguish between healthy and unhealthy breasts with 93% accuracy while determining the size and the size with location with an accuracy of 91% and 74%, respectively.

An In Silico Analysis Identified Members of the Pleckstrin Homology-Like Domain, Family B (PHLDB family) as Potential Prognostic and Predictive Biomarkers of Treatment Response in Breast Cancer Patients.

Breast cancer is the leading cause of morbidity and mortality in women worldwide. This malignant neoplasm can be classified into four clinically relevant subtypes according to the expression of a number of biomarkers. However, these tumors show considerable intratumoral heterogeneity and multidrug resistance. Members of the pleckstrin homology-like domain, family B (PHLDB) play a critical role in the regulation of p53 and AKT signaling pathways, important for cancer and cellular metabolism. The present study was performed to evaluate the expression pattern of PHLDB family members in breast cancer and its potential prognostic and predictive value for therapeutic response using bioinformatics tools. This in silico analysis was performed using several online repositories, including UALCAN, GEPIA2, bc-GenExMiner, KM Plotter, PrognoScan and ROC Plotter. PHLDB family genes were found to be differentially expressed in tumor samples when compared to healthy breast tissue samples. Furthermore, epigenetic regulation may be one of the regulatory mechanisms for the expression of these markers. The PHLDB family of genes proved to be potential markers for predicting the development of lymph node metastasis (p<0.0001) and poor clinical outcome. All members of the PHLDB family were significantly correlated with hormone receptors. High levels of PHLDBs expression were associated with worse overall survival and recurrence-free survival in breast cancer patients. Finally, our data demonstrate that members of the PHLDB family can be promising markers in the stratification of patients who may or may not respond to different available therapies. Our cumulative results demonstrate that PHLDB family members may be promising biomarkers for predicting prognosis and therapeutic response in breast cancer patients.

Tri-Compartmental Restriction Spectrum Imaging Breast Model Distinguishes Malignant Lesions from Benign Lesions and Healthy Tissue on Diffusion-Weighted Imaging.

Simple SummaryWomen at high risk for breast cancer are regularly screened using contrast-enhanced magnetic resonance imaging (MRI) to identify potential malignancy. Diffusion-weighted MRI (DW-MRI) is a non-contrast technique that measures the differential movement of water molecules in tissues and is sensitive to cancer cells. In this study, we use a multi-exponential advanced DW-MRI model called restriction spectrum imaging (RSI) to characterize the diffusion characteristics of malignant lesions, benign lesions, and healthy breast tissue to help differentiate benign from malignant disease. In a cohort of patients, we show that cancer exhibits more restricted diffusion compared to benign breast lesions and healthy tissue, whereas benign and healthy tissue are not different from each other.Diffusion-weighted MRI (DW-MRI) offers a potential adjunct to dynamic contrast-enhanced MRI to discriminate benign from malignant breast lesions by yielding quantitative information about tissue microstructure. Multi-component modeling of the DW-MRI signal over an extended b-value range (up to 3000 s/mm2) theoretically isolates the slowly diffusing (restricted) water component in tissues. Previously, a three-component restriction spectrum imaging (RSI) model demonstrated the ability to distinguish malignant lesions from healthy breast tissue. We further evaluated the utility of this three-component model to differentiate malignant from benign lesions and healthy tissue in 12 patients with known malignancy and synchronous pathology-proven benign lesions. The signal contributions from three distinct diffusion compartments were measured to generate parametric maps corresponding to diffusivity on a voxel-wise basis. The three-component model discriminated malignant from benign and healthy tissue, particularly using the restricted diffusion C1 compartment and product of the restricted and intermediate diffusion compartments (C1 and C2). However, benign lesions and healthy tissue did not significantly differ in diffusion characteristics. Quantitative discrimination of these three tissue types (malignant, benign, and healthy) in non-pre-defined lesions may enhance the clinical utility of DW-MRI in reducing excessive biopsies and aiding in surveillance and surgical evaluation without repeated exposure to gadolinium contrast.

Healthy Eating and Mortality among Breast Cancer Survivors: A Systematic Review and Meta-Analysis of Cohort Studies.

This systematic review examined the effect of diet quality, defined as adherence to healthy dietary recommendations, on all-cause and breast cancer-specific mortality. Web of Science, Medline, CINAHL, and PsycINFO databases were searched to identify eligible studies published by May 2021. We used a random-effects model meta-analysis in two different approaches to estimate pooled hazard ratio (HR) and 95% confidence interval (CI) for highest and lowest categories of diet quality: (1) each dietary quality index as the unit of analysis and (2) cohort as the unit of analysis. Heterogeneity was examined using Cochran’s Q test and inconsistency I2 statistics. The risk of bias was assessed by the Newcastle–Ottawa Scale for cohort studies, and the quality of evidence was investigated by the GRADE tool. The analysis included 11 publications from eight cohorts, including data from 27,346 survivors and seven dietary indices. Both approaches yielded a similar effect size, but cohort-based analysis had a wider CI. Pre-diagnosis diet quality was not associated with both outcomes. However, better post-diagnosis diet quality significantly reduced all-cause mortality by 21% (HR = 0.79, 95% CI = 0.70, 0.89, I2 = 16.83%, n = 7) and marginally reduced breast cancer-specific mortality by 15% (HR = 0.85, 95% CI = 0.62, 1.18, I2 = 57.4%, n = 7). Subgroup analysis showed that adhering to the Diet Approaches to Stop Hypertension and Chinese Food Pagoda guidelines could reduce breast cancer-specific mortality. Such reduction could be larger for older people, physically fit individuals, and women with estrogen receptor-positive, progesterone receptor-negative, or human epidermal growth factor receptor 2-positive tumors. The risk of bias in the selected studies was low, and the quality of evidence for the identified associations was low or very low due to imprecision of effect estimation, inconsistent results, and publication bias. More research is needed to precisely estimate the effect of diet quality on mortality. Healthcare providers can encourage breast cancer survivors to comply with healthy dietary recommendations to improve overall health. (Funding: University of Central Florida Office of Undergraduate Research, Registration: PROSPERO-CRD42021260135).

In silico analysis of expression and DNA methylation profiles of NLRP13 inflammasome in tumor cells

BackgroundNOD-like receptor proteins (NLRs) are essential innate pattern recognition receptors that play fundamental roles in inflammation, and they have been associated with many cancers. NLRP13 (NLR Family Pyrin Domain Containing 13) belongs to the NLRP protein family, but its biological function is not as well defined as other related proteins in this family. Methods and resultsThe expression profile and CpG-aggregated methylation status of the NLRP13 gene were determined using GENT2 and TCGA online platforms. Samples from patients with different tumor stages were used to evaluate the promoter methylation status of NLRP13 in cancer versus normal tissues in a tumor stage-dependent manner by using UALCAN server. We further studied NLRP13 expression in response to inflammatory and hypomethylating agents in two distinct cancer cell lines, HT-29 (human colon adenocarcinoma) and C6 (rat glioma cells), following the treatment with LPS and/or a hypomethylating agent, decitabine, to reveal NLRP13 methylation profiles in these cell lines. ConclusionsUsing bioinformatics tools, we found that NLRP13 expression is higher in healthy breast, head and neck, prostate tissues compared to tumor tissues, whereas its expression is significantly higher in colorectal adenocarcinoma tissue than in normal colon tissue. We also found that NLRP13 gene is mostly hypomethylated in cancer tissues compared to normal tissues, and its high expression is linked with worse overall survival depending on cancer type. Based on data from UALCAN database, NLRP13 promoter methylation is similarly decreased in many cancer patients depending on individual cancer stages. Protein-protein interactions (PPIs) data indicated that NLRP13 showed a confidence interaction with MEFV, NLRC4 and CASP1 proteins, and gene ontology terms associated with functional partners of NLRP13 were also identified in the current study. Our results further showed that NLRP13 expression is regulated by DNA methylation, and treatment with decitabine might effect NLRP13-mediated inflammatory responses in cancer since LPS-induced pro-inflammatory responses specified by IL-1β, IL-18 and caspase-1 levels change in certain cases in cancer cell lines following the pretreatment with this epigenetic modulator. Our study is the first to provide evidence for the differential expression and epigenetic regulation of the NLPR13 in multiple cancer types.

Abstract 282: Breast cancer microphysiological systems based on human breast tissue

Abstract Breast cancer is a heterogeneous disease that accumulates signaling cues from cell intrinsic and extrinsic factors. Many of these cues are derived from the tumor microenvironment (TME). The TME modulates cellular proliferation, survival, and resistance to therapy in breast cancer. The TME is a bio-mechanical and biochemical reservoir for extracellular matrix and signaling factors (cytokines, growth factors, lipids, and hormones). In addition, the TME houses a diverse array of cell populations including adipocytes, vasculature, stem cells, immune cells, and cancer cells. Currently models do not reprpoduce the TME in vitro. The goal of this study was to develop a complex and dynamic micro-physiological 3D tumor model for pre-clinical studies using the native human breast environment. We developed a technique to maintain healthy human breast tissue (HBT) alive ex vivo for up to 8 weeks in the presence of breast cancer (BC) cells creating a breast cancer micro-physiological system (BC-MPS). In this study, we demonstrate alterations to lipid accumulation, matrix fiber remodeling, and cytokine profile in our BC-MPS system. We used RNA sequencing to detect breast cancer cell line signatures retained in the BC-MPS system. Results demonstrate retention of the breast cancer cell line transcriptome for a minimum of 14 days in vitro. In addition we identify alterations to genes associated with the TME, specifically the extracellular matrix and metabolism. Our novel model will allow for 1) real-time examination of BC-HBT interactions and 2) the isolation of BC-specific factors vs. patient-specific factors on the development and progression of BC. Citation Format: Frank H. Lau, C. Ethan Byrne, Loren Brown, Jake Fontenot, Rafael P. Tiongco, Rakesh R. Gurrala, Jonathan Cuccia, Andrew DiNardo, Matthew Burow, Elizabeth C. Martin. Breast cancer microphysiological systems based on human 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 282.