Non-invasive Breast Cancer Research Articles

Empirical wavelet transform based photoacoustic spectral response technique for assessment of ex-vivo breast biopsy tissues

Empirical wavelet transform (EWT) based Photoacoustic spectral response technique is proposed for quantitative assessment of human breast masses (e.g. normal, benign and malignant) based on tissue mechano-biological property. EWT derived parameters such as spectral magnitude and total energy of mono frequency components were utilised for quantitative analysis. Spectral magnitude of second frequency component clearly proves proliferation of epithelial cells in malignant tissues compared to normal. In addition, fibroadenoma illustrates identical energy values of both mono-components which indicates identical proliferation of epithelial and stromal cells. Subsequently, EWT features are proposed that can differentiate the breast masses with 97.5% efficiency, a significant improvement in non-invasive breast cancer diagnosis which also highly correlates with standard histopathology.

Abstract P3-12-13: Comparison of accelerated partial breast irradiation (APBI) with whole breast irradiation (WBI) using 3D conformal external beam radiation therapy (3D CRT)

Abstract Purpose: To compare accelerated partial breast irradiation with whole breast irradiation in post breast conservation surgery (BCS) women with breast cancer. Material and methods: Women >35 years of age with invasive or noninvasive breast cancer ≤4 cm treated by BCS were randomized to 3D CRT APBI (34 Gy in 10 fractions given twice daily) or WBI (40 Gy in 16 fractions given once daily ± boost irradiation).The primary outcome was ipsilateral breast tumour recurrence and important secondary outcomes were adverse cosmetic outcome (fair or poor on a HARVARD/NSABP/RTOG breast cosmesis grading scale) and toxicity using the RTOG scores and LENT SOMA scale. Radiation toxicities and cosmesis was assessed directly by radiation oncologist during radiotherapy and in follow up.Patient and tumour characteristics, locoregional recurrence and distant metastases rates were compared using Fisher's exact tests.All statistical tests were two sided p values less than 0.05 were considered statistically significant. Results: Between June 2011 and December 2015, 133 women were randomized to 3D CRT APBI or WBI. Patient characteristics were balanced between two arms. There was an increased rate of acute grade 2 dermatitis in WBI arm(p=0.33). Median follow up was 60 months(range 9-84 months). Grades 1 and 2 late radiation toxicities were higher in the WBI arm compared to the APBI arm; pigmentation, fibrosis and breast shrinkage were significantly more in the WBI arm. Grades 3 and 4 late toxicity was not seen in any of the treatment arms. Adverse cosmesis at last follow up was significantly higher in patients treated with WBI, 32% as compared to 6% with APBI (p=<0.001). Local recurrence with APBI was 3% as compared to 1.5% with WBI and distant metastasis rate was high in WBI arm as compared with APBI, 5.9% vs 3% respectively; both not statisticaly significant. Table 1.Patient and tumor characteristicsCharacteristicsAPBI(n=65)WBI(n=68)p-valueMean age(range)50(36-75)(36-75)50(37-67)(37-67)0.13T- Stage T134(52)37(54) T228(43)28(41)0.96T33(5)3(4) N stage N057(88)59(87) N17(11)28(41)1.00N21(1)1(1) Grade 114(22)12(18) 237(56)40(59)0.5314(22)16(23) Surgical margins Negative60(92)61(90)0.79Positive5(8)7(10) LVI No56(86)55(81)0.49Yes9(14)13(19) ER Positive42(65)45(66) Negative19(29)21(31)1.00Unknown5(6)2(3) PR Positive38(58)39(57) Negative23(35)25(37)1.0Unknown4(7)4(6) Her2-neu Positive7(11)6(9)0.78Chemotherapy Yes35(54)52(76)0.78No30(46)16(24) Hormone therapy Yes49(75)49(72)0.78No16(25)19(28) Trastuzumab Yes3(43)3(50)1.00No4(57)3(50) Conclusion: In women with BCS, APBI was associated with better cosmetic outcome and late radiation toxicities compared to WBI. Local recurrence and distant metastasis was comparable in both the arms. Citation Format: Yadav BS, L S, Sharma SC, Singh R, Dahiya D. Comparison of accelerated partial breast irradiation (APBI) with whole breast irradiation (WBI) using 3D conformal external beam radiation therapy (3D CRT) [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P3-12-13.

Abstract PD8-08: Development of humanized immune DCIS models using patient peripheral blood derived hematopoietic stem cells (CD34+)

Abstract Background: Ductal carcinoma in situ (DCIS) is the most common form of non-invasive breast cancer. To accurately study the natural progression of DCIS lesions in mice, we devised the mouse-intraductal (MIND) animal model, which involves intraductal injection of human DCIS epithelial cells into the mammary ducts of immunocompromised mice. To improve the translational application of the MIND model, we aimed to mimic the natural microenvironment of human DCIS with patient-derived immune cells and assess the role of engrafted immune cells on human DCIS progression. In order to achieve successful engraftment of the entire immune system in mice, we utilized MISTRG mice. These mice were developed by Rongvaux et al., on an immunodeficient (Rag2-/-IL2rγ-/-) background. The genes encoding human M-CSF (M), human IL-3 (I), SIRP1α (S), human thrombopoietin (TPO)(TR), and GM-CSF (G) were knocked into their respective mouse loci. As such, MISTRG mice are highly permissive for human hematopoiesis, supporting the development and function of lymphocytes, monocytes, and natural killer (NK) cells. In contrast, previous studies have used the humanized CD34+ NOD-SCID IL2rγ-/- mice (CD34+NSG), which are unable to support myeloid cell differentiation due to lack of expression of human-specific cytokines. Moreover, prior xenograft studies in the CD34+NSG mice have not used immune cells derived from the same patient as the tumor. Results: Human CD34+ cells derived from patients' peripheral blood were expanded ex vivo ˜100-fold using a novel formulation of culture medium. Transplantation of ex vivo expanded CD34+ cells via tail vein injection of MISTRG mice resulted in the successful engraftment of human immune cells as early as 4 weeks following injection. Successful engraftment was confirmed by flow cytometry using human specific antibodies that recognize human leukocytes (anti-CD45), T cells (anti-CD3), B cells (anti-CD20), and myeloid cells (anti-CD33) in spleen, bone marrow, and peripheral blood of MISTRG mice. Once engraftment was confirmed, DCIS epithelial cells from the same DCIS patients or DCIS cell lines were injected intraductally. Recruitment of patient-derived immune cells to the DCIS lesions was confirmed by immunofluorescence using human-specific antibodies that recognize neutrophils (anti-myeloperoxidase), macrophages (anti-CD68), M2-polarized macrophages (anti-c-MAF), natural killer cells (anti-CD56), dendritic cells (anti-CD21), T cells (anti-CD3) and B cells (anti-CD20). Conclusion: This model represents the first to enable the study of mechanisms of DCIS progression in a manner that fully represents the heterogeneity of human disease, including the influence of the patients' own immune cells on DCIS progression. Citation Format: Behbod F, Harper H, Hansford H, Limback D, Hong Y, Elsarraj H, Ricci LR, Fan F, Tawfik O, May L, Cusick T, Inciardi M, Redick M, Gatewood J, Winblad O, Fields TA, Fabian C, Godwin AK, Fields PE, Meierotto R, Perry J. Development of humanized immune DCIS models using patient peripheral blood derived hematopoietic stem cells (CD34+) [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr PD8-08.

Fibril bending stiffness of 3D collagen matrices instructs spreading and clustering of invasive and non-invasive breast cancer cells

Extracellular matrix stiffening of breast tissues has been clinically correlated with malignant transformation and poor prognosis. An increase of collagen fibril diameter and lysyl-oxidase mediated crosslinking has been observed in advanced tumor stages. Many current reports suggest that the local mechanical properties of single fibrillar components dominantly regulate cancer cell behavior. Here, we demonstrate by an independent control of fibril diameter and intrafibrillar crosslinking of three-dimensional (3D) collagen matrices that fibril bending stiffness instructs cell behavior of invasive and non-invasive breast cancer cells. Two types of collagen matrices with fibril diameter of either 650 nm or 800 nm at a similar pore size of 10 μm were reconstituted and further modified with the zero-length crosslinker 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide (EDC) at concentrations of 0, 20, 100 and 500 mM. This approach yields two sets of collagen matrices with overlapping variation of matrix elasticity. With these matrices we could prove the common assumption that matrix elasticity of collagen networks is bending dominated with a linear dependence on fibril bending stiffness. We derive that the measured variation of matrix elasticity is directly correlated to the variation of fibril bending stiffness, being independently controlled either by fibril diameter or by intrafibrillar crosslinking. We use these defined matrices to demonstrate that the adjustment of fibril bending stiffness allows to instruct the behavior of two different breast cancer cell lines, invasive MDA-MB-231 (human breast carcinoma) and non-invasive MCF-7 cells (human breast adenocarcinoma). Invasiveness and spreading of invasive MDA-MB-231 cells as well as clustering of non-invasive MCF-7 cells is thereby investigated over a broad parameter range. Our results demonstrate and quantify the direct dependence of cancer cell phenotypes on the matrix mechanical properties on the scale of single fibrils.

Variation in the Use of Boost Irradiation in Breast-Conserving Therapy in the Netherlands: The Effect of a National Guideline and Cofounding Factors

AimsTo determine the variation in radiation therapy boost use in a nationwide study following adjustment of a national guideline in 2011, as well as to address the relationship to patient, tumour and radiation therapy institutional factors. Materials and methodsAll invasive breast cancers and non-invasive breast cancers (ductal carcinoma in situ; DCIS) that received external whole-breast radiation between 2011 and 2016 were selected from the Netherlands Cancer Registry. Box plots were used to evaluate variation over time and logistic regression was carried out to address other factors influencing the variation. Funnel plots were constructed, with unadjusted and adjusted data for patient and tumour factors significantly affecting the use of a boost. ResultsFor breast cancer patients (n = 45,207), the proportion receiving a boost and its range decreased over the years from 37.3–92.7% in 2011 to 28.3–65.4% in 2016. This trend was not observed in DCIS patients (n = 6,844). Young age, large tumours, high grade and the absence of tumour-free resection margins were associated with boost use for both breast cancer and DCIS. For breast cancer, triple-negative tumour subtype and metastatic lymph node involvement were also associated with boost use. Institutional factors did not influence the use of a boost and institutional variation remained substantial after case-mix adjustments. ConclusionFollowing adjustment of a nationwide implemented guideline, variation in radiation therapy boost use decreased in patients with breast cancer but not in patients with DCIS. Several tumour and patient characteristics were associated with boost use. Substantial institutional variation could not be explained by differences in patient, tumour or predefined institutional characteristics.

Breast cancer diagnosis in a specialised breast clinic: Are cancers detected by ultrasound alone less aggressive?

The aim of this study was to determine the relationship between the way breast cancer is diagnosed and its prognostic features. We studied new primary invasive and non-invasive breast cancers (670) diagnosed between 2013 and 2015 where detection included at least clinical examination, mammography and breast ultrasound (BUS). The cancers were classified into six Diagnostic Groups according to the results of each modality. Overall, 79% of cancers were positive on mammography, but another 20% were diagnosed on BUS after a negative mammogram. The largest group (37.6% of cases), had all three modalities positive (Group 1). BUS was the only modality positive in 14.6% (Group 4). Mammography alone was positive in 21.2%, of which 73.9% were ductal carcinoma insitu (DCIS). Invasive lobular carcinoma comprised 9.6% of the groups where mammography was positive, but 16.5% of the groups where mammography was negative and BUS positive. Dense breast tissue was more common in the groups where mammography was negative and BUS positive. Invasive cancers comprised 82.7% of Group 4 and 95.2% of Group 1. For those in Group 4, the average size (10.2mm) and the percentages with lymphovascular invasion (11.1%), lymph node involvement (17.3%) and poor differentiation (12.3%) were less than half the corresponding figures for Group 1 (27.3mm, 35%, 44%, 44%). This study illustrates the complementary benefits of mammography and BUS, especially where breast density is high. Tumours diagnosed by BUS alone had better prognostic features in terms of size, lymphovascular invasion, lymph node status, differentiation and hormone receptors, compared with cancers where all three modalities were positive.

In Situ Detection of Plasma Exosomal MicroRNA-1246 for Breast Cancer Diagnostics by a Au Nanoflare Probe.

Breast cancer is the second cause of cancer mortality in women globally. Early detection, treatment, and metastasis monitoring are of great importance to favorable prognosis. Although conventional diagnostic methods, such as breast X-ray mammography and image positioning biopsy, are accurate, they could cause radioactive or invasive damage to patients. Liquid biopsy as a noninvasive method is convenient for repeated sampling in clinical cancer prognostic, metastatic evaluation, and relapse monitoring. MicroRNAs encased in exosomes circulating in biofluids are promising candidate cancer biomarkers because of their cancer-specific expression profiles. Here, we report an in situ detection of microRNA-1246 (miR-1246) in human plasma exosomes as breast cancer biomarker by a nucleic acid functionalized Au nanoflare probe. Needing neither time-consuming and costly isolation of exosomes from the plasma sample nor transfection means, the Au nanoflare probe can directly enter the plasma exosomes to generate fluorescent signal quantitatively by specifically targeting miR-1246. Only 40 μL of plasma is needed to incubate 4 h with the probe, giving signal sensitive enough to distinguish samples of breast cancer to normal control. Using plasma miR-1246 level detected by our assay as a marker, we differentiated 46 breast cancer patients from 28 healthy controls with 100% sensitivity and 92.9% specificity at the best cutoff. This simple, accurate, sensitive, and cost-effective liquid biopsy by the Au nanoflare probe is potent to be developed as a noninvasive breast cancer diagnostic assay for clinical adaption.

The Influence of Patient Exposure to Breast Reconstruction Approaches and Education on Patient Choices in Breast Cancer Treatment.

BackgroundThe landscape of surgical and medical management and patient choices for breast cancer treatment changes as breast reconstruction and oncoplastic approaches improve and diversify. Increased access to breast reconstruction, in addition to patient education, influences the breast cancer patient. Therefore, the examination of the possible impact of reconstructive surgery on all stages of the breast cancer management per se seemed timely.MethodsPlastic surgery consults were arranged for 520 new patients diagnosed with breast cancer (2012–2016) including patients with noninvasive breast cancer but at high risk of further cancer development. To test the plastic surgery impact on patient choices regarding the management of the cancer, a subset of 90 patients was identified to test the plastic surgery impact on patient choices. These patients were referred to plastic surgery, following the first round of consultations by surgical and medical oncologists with only the preliminary oncological management plan defined. After a plastic surgery consultation, but prior to finalization of the overall oncological management plan, they were surveyed on the subject of modification of their personal choices and requests pertaining to their cancer management.ResultsIn this subset of 90 patients 40 (44%) returned to their surgical or medical oncologist considering changes of the primary management plan after their plastic surgery consultation. Twenty-six (28%) ultimately altered their plan, and the following patient-driven changes were made: mastectomy as opposed to lumpectomy (18 patients [20%]), contralateral prophylactic mastectomy (11 patients [12%]), nipple/areola removal as opposed to nipple/areola sparing suggested by the oncologists (5 patients [6%]), oncoplastic breast reduction as part of lumpectomy (5 patients [6%]), and other modifications (3 patients [3%]).ConclusionsDecisions for altering the preliminary oncologic plan or choosing a specific alternative (eg, lumpectomy plus radiation vs mastectomy) resulted from patient education on (1) reconstructive options, (2) aesthetic pitfalls and results. and (3) their interfacing with the oncological outcomes. Ultimately, plastic surgeons influence the multispecialty breast cancer management and patient decision-making process. Therefore, oncological literacy for plastic surgeons is essential to provide state-of-the-art breast cancer care and avoidance of suboptimal patient decisions.

Oncosmart: A Private Initiative to Improve Breast Cancer Care in Costa Rica

Purpose Founded in 2010, Hospital Metropolitano is the fastest growing health care system in Costa Rica with a network of two hospitals, 27 clinics, and more than 90,000 patients enrolled in its health plan, Medismart. Given the challenges faced by the national public health care system in providing cancer care, the Oncosmart program was launched in November 2016 with the goal of providing population-based cancer screening, diagnostics, cancer treatment, and survivorship care. The program has no restrictions on preexisting conditions and has a monthly membership cost of $14 USD per patient. Here, we summarize our breast cancer program results during the first 16 months of implementation. Methods We performed a retrospective analysis that evaluated data from our radiology department, cancer center, and survivorship program. We determined the number of patients with breast cancer who were diagnosed and/or received care in our health care system from November 2016 to March 2018. Results During this period analyzed, 5,687 mammograms were performed at our health care system and 215 patients were reported to have a Breast Imaging Reporting and Data System 4 to 5 requiring a breast biopsy. Of these, 24% were diagnosed with breast cancer, 36% had a nonmalignant breast condition, and 41% were false positives. Among patients with newly diagnosed breast cancer, 11% had noninvasive breast cancer and 89% had invasive disease. Twenty-seven percent of all breast cancer diagnosis occurred in women younger than age 50 years and two patients were diagnosed with a BRCA1/2 mutation. All biopsies performed at our program were reported within 30 days from abnormal imaging. During this period, we performed 53 breast cancer surgeries and provided chemotherapy and/or endocrine therapy to 32 patients, and among all patients who followed-up in the survivorship clinic, 22 followed-up for breast cancer survivorship. Conclusion Oncosmart is a low-cost and effective private initiative that improves access to breast cancer screening and breast cancer care in the middle-income country of Costa Rica. On the basis of national statistics, after 16 months of implementation, approximately 4% to 5% of all patients with breast cancer in the country are diagnosed within our program. AUTHORS' DISCLOSURES OF POTENTIAL CONFLICTS OF INTEREST The following represents disclosure information provided by authors of this manuscript. All relationships are considered compensated. Relationships are self-held unless noted. I = Immediate Family Member, Inst = My Institution. Relationships may not relate to the subject matter of this manuscript. For more information about ASCO's conflict of interest policy, please refer to www.asco.org/rwc or ascopubs.org/jco/site/ifc . Andres Wiernik Consulting or Advisory Role: Grupo Montecristo Leonardo Lami Employment: Roche, Genetech (I) Travel, Accommodations, Expenses: Roche/Genetech (I) Marissa Durman Consulting or Advisory Role: Grupo Montecristo

Rate of radial scars by core biopsy and upgrading to malignancy or high-risk lesions before and after introduction of digital breast tomosynthesis.

Radial scars (RS) commonly present mammographically as architectural distortions, but these lesions may be associated with non-invasive and invasive breast cancer. Digital breast tomosynthesis (DBT) has resulted in higher detection rates of architectural distortion particularly in patients with dense breast tissue. We hypothesized that rates of clinically relevant lesions confirmed surgically would be lower in patients who received DBT imaging compared with those who received standard digital breast imaging. We performed a retrospective review of 223 patients diagnosed with pure RS by core biopsy and surgical excision before and after DBT was introduced. The rate of upgrading to malignancy or high-risk lesion was evaluated. Demographics, biopsy type, and histologic data were analyzed. Univariable logistic regression analysis was used to identify variables that may be associated with upgrading. The rate of identifying RS increased from 0.04-.13% (P < 0.0001) with DBT imaging. The upgrade rate on surgical specimen to invasive or non-invasive cancer was similar before and after DBT; 6% versus 3%, as were findings of a high-risk lesion; 12% versus 22%. No predictive factors were identified for patients upgraded to malignant neoplasms or high-risk lesions. The likelihood of identifying RS has increased with DBT imaging, but rates of upgrading to a malignant neoplasm or high-risk lesion were similar to those before DBT. Although the rate of upgrading to malignancy after DBT was low, an excisional biopsy should be considered as 22% of patients were upgraded to high-risk lesions. These patients are candidates for chemoprevention and/or high-risk surveillance.

Loss of mdig expression enhances DNA and histone methylation and metastasis of aggressive breast cancer

We previously reported that expression of an environmentally induced gene, mineral dust-induced gene (mdig), predicts overall survival in breast cancer patients. In the present report, we further demonstrate the differential roles of mdig between earlier- and later-stage breast cancers. In noncancerous breast, mdig is a proliferation factor for cell growth and cell motility. In breast cancer, however, higher levels of mdig negatively regulate the migration and invasion of cancer cells. Assessment of global DNA methylation, chromatin accessibility and H3K9me3 heterochromatin signature suggests that silencing mdig enhances DNA and histone methylation. Through immunostaining and data mining, we found that mdig is significantly upregulated in noninvasive and/or earlier-stage breast cancers. In contrast, in triple-negative and other invasive breast cancers, diminished mdig expression was noted, indicating that the loss of mdig expression could be an important feature of aggressive breast cancers. Taken together, our data suggest that mdig is a new biomarker that likely promotes tumor growth in the early stages of breast cancer while acting as a tumor suppressor to inhibit invasion and metastasis in later-stage tumors.

TNFAIP3 is required for FGFR1 activation-promoted proliferation and tumorigenesis of premalignant DCIS.COM human mammary epithelial cells

BackgroundAlthough ductal carcinoma in situ (DCIS) is a non-invasive breast cancer, many DCIS lesions may progress to invasive cancer and the genes and pathways responsible for its progression are largely unknown. FGFR1 plays an important role in cell proliferation, differentiation and carcinogenesis. The purpose of this study is to examine the roles of FGFR1 signaling in gene expression, cell proliferation, tumor growth and progression in a non-invasive DCIS model.MethodsDCIS.COM cells were transfected with an empty vector to generate DCIS-Ctrl cells. DCIS-iFGFR1 cells were transfected with an AP20187-inducible iFGFR1 vector to generate DCIS-iFGFR1 cells. iFGFR1 consists of the v-Src myristoylation membrane-targeting sequence, FGFR1 cytoplasmic domain and the AP20187-inducible FKBP12 dimerization domain, which simulates FGFR1 signaling. The CRISPR/Cas9 system was employed to knockout ERK1, ERK2 or TNFAIP3 in DCIS-iFGFR1 cells. Established cell lines were treated with/without AP20187 and with/without FGFR1, MEK, or ERK1/2 inhibitor. The effects of these treatments were determined by Western blot, RNA-Seq, real-time RT-PCR, cell proliferation, mammosphere growth, xenograft tumor growth, and tumor histopathological assays.ResultsActivation of iFGFR1 signaling in DCIS-iFGFR1 cells enhanced ERK1/2 activities, induced partial epithelial-to-mesenchymal transition (EMT) and increased cell proliferation. Activation of iFGFR1 signaling promoted DCIS growth and progression to invasive cancer derived from DCIS-iFGFR1 cells in mice. Activation of iFGFR1 signaling also altered expression levels of 946 genes involved in cell proliferation, migration, cancer pathways, and other molecular and cellular functions. TNFAIP3, a ubiquitin-editing enzyme, is upregulated by iFGFR1 signaling in a FGFR1 kinase activity and in an ERK2-dependent manner. Importantly, TNFAIP3 knockout not only inhibited the AP20187-induced proliferation and tumor growth of DCIS-iFGFR1 cells, but also further reduced baseline proliferation and tumor growth of DCIS-iFGFR1 cells without AP20187 treatment.ConclusionsActivation of iFGFR1 promotes ERK1/2 activity, EMT, cell proliferation, tumor growth, DCIS progression to invasive cancer, and altered the gene expression profile of DCIS-iFGFR1 cells. Activation of iFGFR1 upregulated TNFAIP3 in an ERK2-dependent manner and TNFAIP3 is required for iFGFR1 activation-promoted DCIS.COM cell proliferation, mammosphere growth, tumor growth and progression. These results suggest that TNFAIP3 may be a potential target for inhibiting DCIS growth and progression promoted by FGFR1 signaling.

TGF-β induces changes in breast cancer cell deformability

Mechanical properties of cells are shown to regulate cell behaviors leading to phenotypic changes that may aid in the development and progression of disease. In this study, we used atomic force microscopy (AFM) indentation with a spherical probe to characterize the elastic and viscoelastic properties of invasive (MDA-MB-231) and noninvasive (MCF-7) breast cancer cells treated with transforming growth factor-β (TGF-β). We also used confocal fluorescence imaging to investigate the sub-membrane cytoskeletal structure of the cells. Results showed significant alterations in moduli of both cell types after 24 h TGF-β treatment which had a context dependent response; moduli for MDA-MB-231 decreased whereas MCF-7 demonstrated stiffening response. Stress relaxation tests showed increased fluid-like nature of MDA-MB-231 following TGF-β treatment and lower fluidity for MCF-7 cells. We also observed significant alterations in the expression and orientation of actin stress fibers with TGF-β treatment which correlated with the changes in cell mechanics. Less invasive MCF-7 cells had a delayed overall increase in cell deformability after 48 h exposure to TGF-β; a similar trend was observed for MDA-MB cells. These changes may be important to facilitate migration, for instance, during metastasis of cancer cells through submicron sized spaces.

Abstract 185: p300 and STAT3 drive YAP-independent mechanotransduction during breast cancer invasion

Abstract 83% of non-invasive breast cancers are diagnosed as ductal carcinoma in situ (DCIS). While some DCIS tumors remain confined in the mammary duct, about half progress into invasive ductal carcinoma (IDC), when carcinoma cells break through the basement membrane (BM) into the type-1 collagen (col-1) rich stroma, a key first step towards metastasis. As 90% of cancer-related deaths are due to metastasis, preventing progression to invasive disease could significantly reduce cancer mortality. However there are no established biomarkers for invasive potential and current diagnostic methods cannot predict which DCIS cases will progress to IDC. Interestingly, increased tissue stiffness correlates with invasion and the transcriptional regulator YAP has been implicated as a mechanotransducer, largely based on 2D culture studies. To identify the drivers of DCIS progression, we encapsulated mammary epithelial cells (MECs) in 3D hydrogels with a range of stiffness and that presented either BM-ligands or col-1 containing microenvironments to the cells. RNA-seq identified the global gene expression changes induced by increased 3D culture stiffness in BM-like environments. 3SEQ analysis of breast cancer patient samples revealed that genes regulated by 3D culture stiffness were upregulated in breast cancer patients, demonstrating the relevance of 3D culture models and suggesting expression of S100A7 as a potential biomarker of breast cancer progression. Interestingly gene expression changes induced by increased 3D stiffness in BM-like environments were distinct from col-1 like environments. Col-1 exposure in stiff gels promotes expression of genes whose protein products remodel the col-1 network including FN1 and LOX. As col-1 remodeling promotes cell dissemination, this suggests that col-1 exposure following BM invasion induces pro-metastatic changes in carcinoma phenotype. Surprisingly, enhanced stiffness induced invasion in MECs independently of YAP activation in both BM-like and col-1 rich 3D hydrogels. Instead, bioinformatic analysis identified transcriptional regulators p300 and STAT3 as mediators of 3D mechanosensing. Inhibition of p300 and STAT3 in stiff 3D BM-like environments and, conversely, overexpression in soft 3D BM-like environments confirm a role during stiffness-induced proliferation. This suggests p300 and STAT3 as possible targets for preventing progression to invasive disease. While genetic alterations initiate transformation, these results reveal the miroenvironment events that initiate breast cancer invasion. Citation Format: Joanna Y. Lee, Jessica Chang, Sungmin Nam, Hong-pyo Lee, Antonia A. Dominguez, Sushama Varma, Lei S. Qi, Robert B. West, Ovijit Chaudhuri. p300 and STAT3 drive YAP-independent mechanotransduction during breast cancer invasion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 185.

Breast Cancer

Invasive breast cancer, the most common nonskin cancer in women in the United States, will be diagnosed in 266,120 In 2018, along with 63,960 new cases of non-invasive (in situ) breast cancer. Incidence and mortality reached a plateau and appear to be dropping in both the United States and parts of western Europe. This decline has been attributed to several factors, such as early detection through the use of screening mammography and appropriate use of systemic adjuvant therapy, as well as decreased use of hormone replacement therapy. However, the global burden of breast cancer remains great, and global breast cancer incidence increased from 641,000 in 1980 to 1,643,000 in 2010, an annual rate of increase of 3.1%. This chapter examines the etiology, epidemiology, prevention, screening, staging, and prognosis of breast cancer. The diagnoses and treatments of the four stages of breast cancer are also included. Figures include algorithms used for the systemic treatment of stage IV breast cancer and hormone therapy for women with stage IV breast cancer. Tables describe selected outcomes from the National Surgical Adjuvant Breast and Bowel Project (NSABP) P-1 and P-2 chemoprevention trials, tamoxifen chemoprevention trials for breast cancer, the TNM staging system and stage groupings for breast cancer, some commonly used adjuvant chemotherapy regimens, an algorithm for suggested treatment for patients with operable breast cancer from the 2011 St. Gallen consensus conference, guidelines for surveillance of asymptomatic early breast cancer survivors from the American Society of Clinical Oncology, and newer agents for metastatic breast cancer commercially available in the United States. This review contains 2 highly rendered figures, 8 tables, and 108 references.

Treatment patterns for unilateral, non-invasive breast cancer in women diagnosed in England: Data from a population-based cohort

Introduction: Population-based studies of non-invasive breast cancer in older women are infrequently reported. We evaluated the management of ductal carcinoma in situ (DCIS), in women aged >=70yrs compared to those aged 50 – 69yrs, as part of the National Audit of Breast Cancer in Older Patients (NABCOP).

Inhibition of breast cancer cell growth by the combination of clofarabine and sulforaphane involves epigenetically mediated CDKN2A upregulation

ABSTRACTMany antineoplastic nucleoside analogue-based combinatorial strategies focused on remodelling aberrant DNA methylation patterns have been developed. The number of studies demonstrate high efficacy of bioactive phytochemicals in support of conventional chemotherapy. Our recent discoveries of the epigenetic effects of clofarabine (2′-deoxyadenosine analogue, antileukaemic drug) and clofarabine-based combinations with dietary bioactive compounds in breast cancer cells led us to look for more DNA methylation targets of these cancer-preventive agents. In the present study, using methylation-sensitive restriction analysis (MSRA) and qPCR, we showed that clofarabine in combination with sulforaphane, a phytochemical from cruciferous vegetables, significantly reactivates DNA methylation-silenced CDKN2A tumour suppressor and inhibits cancer cell growth at a non-invasive breast cancer stage.

Distinct Effect of Invasive and Non‐invasive Breast Cancer Cells on Vasculogenic Capacity of Endothelial Progenitors

Blood vessel formation plays an essential role in tumor growth and metastasis. Cancer cells secrete various pro‐angiogenic factors that activate preexisting vascular endothelial cells as well as induce recruitment of bone‐marrow‐derived endothelial progenitors; endothelial colony forming cells (ECFC). The interactions between cancer cells and endothelial cells have been largely investigated, but it has been rarely studied whether the vasculogenic capacity of ECFC are differently affected by invasive and non‐invasive phenotype of the cancer cells. Thus, we investigated to determine the vasculogenic potentials of invasive and non‐invasive breast cancer cells. MDA‐MB‐231 and MCF7 were applied as invasive and non‐invasive breast cancer cells. Bone marrow‐derived ECFC were isolated from human peripheral blood. Tubular structure number and length as well as spheroid sprout number and length formed by ECFC was increased when treated with MDA‐MB‐231‐derived conditioned medium (CM) compared with MCF7‐derived CM. Migration of ECFC was also increased by treatment of MDA‐MB‐231‐derived CM compared to MCF7. To mimic tumor microenvironment, ECFC were directly co‐cultured with cancer cells. Tube formation of ECFC was enhanced under the co‐culture condition with MDA‐MB‐231 compared to ECFC alone control. Interestingly, ECFC co‐cultured with MCF7 significantly decreased tube formation. Our results suggest that MDA‐MB‐231 and MCF7 promote ECFC vasculogenesis in different manners, which may be the important determinants to characterize invasive and non‐invasive phenotype of the cancer cells.Support or Funding InformationThis research was supported by Priority Research Centers Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2016r1a6a1a03007648), and supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT (2017R1A2B4005463).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

A Blockade of IL‐6 Released from Human Breast Carcinoma MDA‐MB‐231 in Presence of NK‐92 Inhibits an Invasion of Breast Carcinoma

A variety of cytokines are existed in tumor microenvironment, which might affect tumorigenesis and metastasis. Recent studies have demonstrated that interleukin(IL)‐6, a pro‐inflammatory cytokine, is significantly present in breast tumor microenvironment. It is suggested to have a critical role in an epithelial to mesenchymal transmission(EMT) process in breast cancer cells. Natural killer (NK) cells are a large innate lymphocyte that has a cytotoxic effect to cancer cells or virus‐infected cells. However, in breast tumor microenvironment, NK cells are shown to have pro‐tumor activity in presence of various inflammatory cytokines and growth factors. We found that IL‐6 is highly produced in coculture of invasive breast cancer cell MDA‐MB‐231 and natural killer(NK)‐92 cell, but is little produced in coculture of non‐invasive breast cancer cell MCF‐7 and NK cells. Subsequently, we found that the expression of p‐stat3, p‐erk, akt and pi3k proteins is statistically up‐regulated in a coculture of MDA‐MB‐231 and NK‐92 compared to in coculture with MCF‐7. A blockade of IL‐6 by anti‐IL‐6 (1ng/ml) or shRNA in invasive breast cancer cell MDA‐MB‐231 reduced a migratory and invasive capability as well as down‐regulated an expression of p‐stat3 protein. Further we examined the inhibitory effect of apigenin on invasion or migration of invasive breast cancer cell MDA‐MB‐231. We found that a blockade of IL‐6 by apigenin in MDA‐MB‐231 mediates the inhibitory effect on migration and invasion of MDA‐MB‐231.Support or Funding Informationthis research was supported by basic science research program through the national research foundation of korea(nrf) funded by the ministry of education(2016R1A6A1A03007648)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Population-based study of factors influencing invasive breast cancer risk after screen-detected ductal carcinoma in situ: First results from the non-invasive breast cancer in England (NINBE) study

Population-based study of factors influencing invasive breast cancer risk after screen-detected ductal carcinoma in situ: First results from the non-invasive breast cancer in England (NINBE) study