Enabling the Study of Metabolism in Breast Cancer Through Collection of Fresh-tissue Biopsies

2019 ◽  
Author(s):  
Keyword(s):  
Breast Cancer ◽  
Fresh Tissue

Feasibility of testing core needle biopsies ex vivo in the ChemoFx assay

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 20073-20073
Author(s):  
J. A. O’Shaughnessy ◽  
F. Holmes ◽  
P. Beitsch ◽  
J. Cunningham ◽  
H. Gallion ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Epithelial Cells ◽  
Primary Breast Cancer ◽  
Ex Vivo ◽  
New Technology ◽  
Percutaneous Biopsy ◽  
Response To Therapy ◽  
Fresh Tissue ◽  
Core Needle ◽  
Precision Therapeutics

20073 Background: Multiple chemotherapy options exist for the treatment of primary breast cancer. While response rates are good, many patients are treated with unnecessary or ineffective chemotherapy. Inadequate treatments are partly due to the lack of accurate predictors of response in individual patients. To predict an individual’s response to therapy, ex vivo chemosensitivity and resistance assays (CSRAs) have long been evaluated, but have been limited by technical difficulties, including the need for large (1–2 gm) amounts of fresh tissue. However, these problems have largely been overcome with new technology. Novel methods used in Precision Therapeutics’ ChemoFx assay allow for testing smaller amounts of tissue (35 mg). The reduced tissue requirement is crucial in the breast cancer setting, as the diagnosis is often made by percutaneous biopsy. The goals of the study were to determine the growth success rate of culturing epithelial cells from breast tissue core needle biopsies and the feasibility of testing the cells in the assay. Methods: A prospective feasibility study involving women with invasive primary breast cancer. One to four core needle biopsy specimens were collected using a 14 gauge needle (est. per patient yield <50 mg) and submitted to Precision Therapeutics. A primary culture of each specimen was established and the ex vivo chemoresponse profiles of each culture were evaluated. Drugs tested included capecitabine, cisplatin, cyclophosphamide, docetaxel, doxorubicin, epirubicin, etoposide, 5-fluorouracil, gemcitabine, irinotecan, paclitaxel, and vinorelbine. Results: 21 of 25 (84%, 95% CI: 68% to 97%) specimens grew successfully; all 21 were tested for chemoresponsiveness with the assay. Of the 4 subjects with unsuccessful ex vivo cultures, 2 had no growth, 1 failed plating for culture, and 1 failed IHC testing due to overgrowth of non-epithelial cells. The average number of drugs tested for each specimen was 7 (range: 1–15). Conclusions: This study demonstrates that core needle biopsies from primary breast tumors can be successfully cultured and tested for chemoresponsiveness using the ChemoFx assay. The ability to perform ex vivo chemoresponse testing on core needle biopsies greatly increases the utility of the assay in adjuvant or neoadjuvant primary breast cancer settings. [Table: see text]

Breast cancer stem cells mutations: A new understanding of intratumoral heterogeneity.

2013 ◽  
Vol 31 (26_suppl) ◽  
pp. 25-25
Author(s):  
Cory Donovan ◽  
Amy Skinner ◽  
Rodney F. Pommier ◽  
Jennifer L. Alabran ◽  
Patrick Muller ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Lymph Node ◽  
Lymph Node Metastases ◽  
Lymph Node Status ◽  
Significant Heterogeneity ◽  
Axillary Lymph ◽  
Fresh Tissue ◽  
Axillary Lymph Node Metastases ◽  
Mutation Status ◽  
Node Metastases

25 Background: Breast cancer has long been recognized as a heterogeneous disease. This has profound implications for diagnosis, treatment and disease recurrence. Oncogenic mutations have been identified in breast cancer cells with stem-like and progenitor properties (BCSC). We have previously reported that BCSC mutations correlated with axillary lymph node metastases. This was even more significant when micrometastatic disease was included. Our hypothesis is that tumor heterogeneity extends to the genetics of BCSC, and that BCSC mutations are better predictors of lymph node status than whole tumor genetics. Methods: BCSC from fresh tissue specimens were matched to their whole tumor specimens. BCSC and whole tumor DNA were sent for PCR-based mutation analysis. Patient data was collected by chart review. Results: Twenty-eight matched BCSC and whole tumor samples were analyzed. PI3K/Akt signaling mutations in PIK3CA, AKT1, HRAS, and MET were identified in BCSC from 10 tumors. In 4 of these, mutations were also identified in the corresponding whole tumor specimens. In 4 patients, mutations were identified in whole tumor samples only. Fourteen tumors had no mutations. Tumor stage, grade, receptor status, and age did not correlate with tumor or BCSC mutation status. In contrast to BCSC mutations, mutation status of the whole tumor did not correlate with micro or macro metastatic disease in the lymph node (p = 0.92). Conclusions: Mutations in BCSC are more predictive of lymph node metastases than mutations identified in the tumors. Thus, PI3K/Akt pathway mutations in tumor precursor cells may have a stronger influence on tumor metastatic potential than mutations identified in whole tumor samples. Whole tumors and BCSC populations demonstrate significant heterogeneity, as mutations identified in BCSC and tumors were not always concordant. Rare BCSC populations must be tested separately as they provide crucial prognostic and treatment information in conjunction with whole tumor genetic analyses.

Quantitative nuclease protection assay (qNPA) for gene expression analysis on breast cancer core biopsies.

2011 ◽  
Vol 29 (27_suppl) ◽  
pp. 49-49 ◽  
Author(s):  
M. C. Evangelist ◽  
J. Snider ◽  
J. Krushkal ◽  
Y. Qu ◽  
A. Kulkarni ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Small Sample Size ◽  
Small Sample ◽  
Ffpe Tissue ◽  
Cancer Tissue ◽  
Breast Cancer Patients ◽  
Fresh Tissue ◽  
Interacting Protein ◽  
Oncogenic Pathways

49 Background: qNPA employs in situ hybridization of detection probes to cross linked mRNA, making it ideal for formalin fixed paraffin embedded (FFPE) tissue. It has been shown to measure gene expression in archived lymphoid and lung cancer tissue. We assessed the feasibility of qNPA to measure differentially expressed genes in pretreatment FFPE core breast biopsies among pathologic responders (pR) and nonresponders (pNR) to preoperative chemotherapy. Methods: We included preoperative breast cancer patients treated at our institution from 2003-09 with FFPE core biopsies. mRNA expression of 170 genes, representing oncogenic pathways or associated with anthracycline and taxane response was measured by qNPA (HTG, Tucson, AZ). Data was normalized to 3 housekeeper genes and average of 3 biologic replicates reported. Seven genes below detection in > 50% samples were excluded. Expression values of 163 unique genes were analyzed for pR vs pNR with dChip software. Empirical FDR was estimated using 1,000 permutations of sample labels. Results: Treatment and response: Sample failure: 6/57 (10%). pR vs pNR did not separate on hierarchical clustering. FLJ12650 and IGFBP2 showed lower expression in pR vs pNR with fold changes of 4.09 and 2.40, respectively (p < 0.01; median FDR: 1/163). FLJ12650 was significant (p < 0.01, median FDR: 0/163) when patients receiving anthracycline ± taxane were analyzed (groups 1-3) and showed a trend (p < 0.05) in group 1 alone. Conclusions: qNPA for limited available FFPE tissue from core biopsies is feasible with acceptable sample failure rates. Small sample size and number of analyzed genes limited definitive conclusion about informative genes in our study. Our FLJ12650 results, a gene coding for membrane Na+/K+ ATPase interacting protein, are consistent with previous findings of overexpression in pNR to anthracyclines + taxanes (Hess et al, JCO 2006, Vol 24; 4236) using fresh tissue. Future qNPA validation of predictive markers, identified by whole transcriptome analysis in a homogenous cohort may provide more definitive results. [Table: see text]

FOXA1 expression in Iraqi women with ER+ breast cancer: A case control study

2020 ◽  
Vol 2 (02) ◽  
Author(s):  
Iman Al-Bedairy ◽  
Mais Shamsa ◽  
Safaa aldeen Salim ◽  
Mohammed Mahdi ◽  
Karam Dawood ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Molecular Subtype ◽  
Estrogen Receptor Α ◽  
Luminal Breast Cancer ◽  
Molecular Characteristics ◽  
Luminal A ◽  
Fresh Tissue ◽  
Iraqi Women ◽  
Foxa1 Expression

Background: Breast cancer is a heterogeneous disease that can be classified into many subtypes according to histopathological and molecular characteristics. Forkhead box protein A1 (FOXA1) is a pioneer factor of estrogen receptor (α-ER)-chromatin binding and function. FOXA1 expression is related to luminal breast cancer with a good prognosis. Objectives: The present study is sought to determine the gene expression of FOXA1 in Iraqi women with ER+ breast cancer from fresh tissue. Methods: Forty-eight fresh malignant breast tissues analyzed by immunohistochemistry assay to choose ER+ sample submitted to RT-qPCR to evaluate FOXA1 gene expression. Results: ER-positive was 72.91% of the total samples, luminal A was the most common molecular subtype with 56.25%. Conclusions: Highly significant FOXA1 gene expression in Iraqi women with breast cancer makes it eligible to be a good predictor or a biomarker for breast cancer.

Nanomechanical profiling of human breast tumors as prognostic marker for breast cancer.

2017 ◽  
Vol 35 (15_suppl) ◽  
pp. 11618-11618
Author(s):  
Rosemarie Anne Burian ◽  
Tobias Appenzeller ◽  
Philipp Oertle ◽  
Christian Raez ◽  
Roderick Y.H. Lim ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Clinical Setting ◽  
Risk Groups ◽  
Breast Cancer Subtypes ◽  
Human Breast ◽  
Fresh Tissue ◽  
Fixed Tissue ◽  
Cancer Subtypes ◽  
Nanomechanical Properties ◽  
Cancer Aggressiveness

11618 Background: Assessment of tumor aggressiveness is crucial when making treatment decisions. Established prognostic markers may be insufficient to stratify cancer patients into treatment relevant risk groups. Emerging evidence indicates mechanical properties of cancer cells and their microenvironment play a vital role in cancer invasion and metastases. Detecting and measuring these nanomechanical changes could be a marker of cancer aggressiveness. Methods: We developed an atomic force microscope (AFM) based method: ARTIDIS (Automated and Reliable Tissue Diagnostics) for measuring nanomechanical properties of human tissue biopsies. These were performed on fresh, non-fixed tissue under physiological conditions. This novel method uses a micro-fabricated 20nm tip indenting and measuring stiffness of thousands of locations within 60-180minutes. This quantitative, biopsy-wide, nanomechanical profile strongly correlates to the tissue's biological composition. Post-AFM this biopsy is analyzed by pathology. We sought to differentiate benign from cancerous lesions based on nanomechanical properties; then link the cancerous nanomechanical profiles prospectively to the clinical outcomes. Results: Our results demonstrate the first AFM based nanomechanical profiling to detect aggressive breast cancer subtypes using fresh tissue in a clinical setting. We have shown that nanomechanical profiles of human breast cancer biopsies display stiffness profiles distinct from surrounding normal tissue. Breast cancer subtypes were distinguishable by their nanomechanical properties only. We have discovered specific nanomechanical profiles of tumor subtypes likely to metastasize. When the primary tumor displayed the same soft nanomechanical profile as adjacent tissue, this was associated with positive nodal status. Conclusions: Our results demontrate nanomechanical profiling is a fast and sensitive method to stratify malignant biopsies into relevant subgroups in a clinical setting. Relative stiffness and distribution values provide a nanomechanical profile indicating cancer aggressiveness. This will help optimize specific cancer diagnosis, orientate therapy choice and support patient follow up.

scholarly journals A New Gene Expression Signature for Triple-Negative Breast Cancer using Frozen Fresh Tissue before Neoadjuvant chemotherapy

2017 ◽  
Vol 23 (1) ◽  
pp. 101-111 ◽  
Author(s):  
Sandra K. Santuario-Facio ◽  
Servando Cardona-Huerta ◽  
Yadira X. Perez-Paramo ◽  
Victor Trevino ◽  
Francisco Hernandez-Cabrera ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Neoadjuvant Chemotherapy ◽  
Triple Negative Breast Cancer ◽  
Triple Negative ◽  
Gene Expression Signature ◽  
Fresh Tissue ◽  
Expression Signature ◽  
New Gene

Touch Imprint Cytology in Tumor Tissue Banks for the Confirmation of Neoplastic Cellularity and for DNA Extraction

2008 ◽  
Vol 132 (6) ◽  
pp. 974-978 ◽  
Author(s):  
Anita Mangia ◽  
Annalisa Chiriatti ◽  
Patrizia Chiarappa ◽  
Maria Angela Incalza ◽  
Giovanni Antonaci ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Dna Extraction ◽  
Lymph Node Status ◽  
Tissue Banks ◽  
Imprint Cytology ◽  
Fresh Tissue ◽  
Tissue Samples ◽  
Touch Imprint Cytology ◽  
Biological Studies ◽  
Κ Statistic

Abstract Context.—Learning the characteristics of frozen tissue samples stored in tumor banks for biological studies remains a problem. Objective.—To assess the use of touch imprint cytology on fresh tissue samples as a rapid and reliable method of determining the presence and quantity of neoplastic cells before freezing. Design.—Touch imprint cytology was performed on 259 specimens of operable breast cancer. Touch imprints were prepared from fresh tissue specimens before freezing samples for storage. Each tumor sample was imprinted on a glass slide and stained with hematoxylin-eosin. Tumor cellularity was quantified as negative, poor, moderate, or rich. Results.—A significant correlation was found between samples with a tumor size greater than 2 cm and high tumor cellularity (P = .03; χ2 test). Furthermore, 35% of ductal tumors showed higher tumor cellularity compared with lobular tumors (P &lt; .001; χ2 test). No association was found between lymph node status and tumor grade. When samples for which more than 2 imprints were available were examined, tumor cellularity among imprints of the same sample showed an overall agreement of 0.67 (P &lt; .001; κ statistic). It was also determined that the higher the cellularity, the higher the agreement. Our data also showed concordance of 0.87 (P &lt; .001; κ statistic) between touch imprint cytology imprints and histologic sections from contiguous tumor. Moreover, 11 randomly selected samples underwent DNA extraction, polymerase chain reaction, and sequencing to verify the feasibility of DNA analyses. We found that DNA from touch imprint cytology was amplifiable and suitable for direct sequencing. Conclusions.—Touch imprint cytology may represent an important step in the quality control of tumor cellularity of breast cancer specimens designed to be stored in tumor biobanks and a valid method for assessing the suitability of such tissue for further biomorphologic and biomolecular applications.

SEM Cold Stage Techniques for the Observation of Vegetative and Floral Apices of Oat (Avena sativa L.) Plants

1977 ◽  
Vol 35 ◽  
pp. 590-591
Author(s):  
B. K. Kirchoff ◽  
L.F. Allard ◽  
W.C. Bigelow
Keyword(s):  
Critical Point ◽  
Avena Sativa ◽  
Substantial Improvement ◽  
Fresh Tissue ◽  
Cold Stage ◽  
Critical Point Drying ◽  
Almost All ◽  
Cell Collapse ◽  
Conductive Paint ◽  
Carbon Based

In attempting to use the SEM to investigate the transition from the vegetative to the floral state in oat (Avena sativa L.) it was discovered that the procedures of fixation and critical point drying (CPD), and fresh tissue examination of the specimens gave unsatisfactory results. In most cases, by using these techniques, cells of the tissue were collapsed or otherwise visibly distorted. Figure 1 shows the results of fixation with 4.5% formaldehyde-gluteraldehyde followed by CPD. Almost all cellular detail has been obscured by the resulting shrinkage distortions. The larger cracks seen on the left of the picture may be due to dissection damage, rather than CPD. The results of observation of fresh tissue are seen in Fig. 2. Although there is a substantial improvement over CPD, some cell collapse still occurs.Due to these difficulties, it was decided to experiment with cold stage techniques. The specimens to be observed were dissected out and attached to the sample stub using a carbon based conductive paint in acetone.

Virus-Like Particles in a Human Breast Cancer: Structural Similarity to Previously Reported Particles

1973 ◽  
Vol 31 ◽  
pp. 378-379
Author(s):  
G. Kasnic ◽  
S. E. Stewart ◽  
C. Urbanski
Keyword(s):  
Breast Cancer ◽  
Biological Activity ◽  
Human Breast Cancer ◽  
Osteogenic Sarcoma ◽  
Human Tumor ◽  
Structural Similarity ◽  
Cell Tumor ◽  
Human Breast ◽  
Human Tumor Cell ◽  
Type Particle

We have reported the maturation of an intracisternal A-type particle in murine plasma cell tumor cultures and three human tumor cell cultures (rhabdomyosarcoma, lung adenocarcinoma, and osteogenic sarcoma) after IUDR-DMSO activation. In all of these studies the A-type particle seems to develop into a form with an electron dense nucleoid, presumably mature, which is also intracisternal. A similar intracisternal A-type particle has been described in leukemic guinea pigs. Although no biological activity has yet been demonstrated for these particles, on morphologic grounds, and by the manner in which they develop within the cell, they may represent members of the same family of viruses.

Further Ultrastructural Observations on Metastatic Nodules of Human Breast Cancer

1968 ◽  
Vol 26 ◽  
pp. 224-225
Author(s):  
John L. Swedo ◽  
R. W. Talley ◽  
John H. L. Watson
Keyword(s):  
Breast Cancer ◽  
Human Breast Cancer ◽  
Estrogen Therapy ◽  
Specimen Preparation ◽  
Human Breast ◽  
Autophagic Vacuoles ◽  
Previous Communication ◽  
Linear Profiles ◽  
Metastatic Nodule

Since the report, which described the ultrastructure of a metastatic nodule of human breast cancer after estrogen therapy, additional ultrastructural observations, including some which are correlative with pertinent findings in the literature concerning mycoplasmas, have been recorded concerning the same subject. Specimen preparation was identical to that in.The mitochondria possessed few cristae, and were deteriorated and vacuolated. They often contained particulates and fibrous structures, sometimes arranged in spindle-shaped bundles, Fig. 1. Another apparent aberration was the occurrence, Fig. 2 (arrows) of linear profiles of what seems to be SER, which lie between layers of RER, and are often recognizably continuous with them.It was noted that the structure of the round bodies, interpreted as within autophagic vacuoles in the previous communication, and of vesicular bodies, described morphologically closely resembled those of some mycoplasmas. Specifically, they simulated or reflected the various stages of replication reported for mycoplasmas grown on solid nutrient. Based on this observation, they are referred to here as “mycoplasma-like” structures, in anticipation of confirmatory evidence from investigations now in progress.

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