Christina Finlayson

Cytokeratin 5 positive cells represent a steroid receptor negative and therapy resistant subpopulation in luminal breast cancers.

A majority of breast cancers are estrogen receptor (ER) positive and have a luminal epithelial phenotype. However, these ER+ tumors often contain heterogeneous subpopulations of ER− tumor cells. We previously identified a population of cytokeratin 5 (CK5) positive cells within ER+ and progesterone receptor positive (PR+) tumors that is both ER−PR− and CD44+, a marker of breast tumor-initiating cells (TICs). These CK5+ cells have properties of TICs in luminal tumor xenografts, and we speculated that they are more resistant to chemo- and anti-ER-targeted therapies than their ER+ neighbors. To test this, we used ER+PR+ T47D and MCF7 breast cancer cells. CK5+ cells had lower proliferative indices than CK5− cells, were less sensitive to 5-fluorouracil and docetaxel, and cultures became enriched for CK5+ cells after treatments. CK5+ cells were less prone to drug-induced apoptosis than CK5− cells. In cells treated with 17β-estradiol (E) plus anti-estrogens tamoxifen or fulvestrant, ER protein levels decreased, and CK5 protein levels increased, compared to controls treated with E alone. In ER+ tumors from patients treated with neoadjuvant endocrine therapies ER gene expression decreased, and CK5 gene expression increased in post compared to pre-treatment tumors. The number of CK5+ cells in tumors also increased in post- compared to pre-treatment tumors. We conclude that an ER−PR−CK5+ subpopulation found in many luminal tumors is resistant to standard endocrine and chemotherapies, relative to the majority ER+PR+CK5− cells. Compounds that effectively target these cells are needed to improve outcome in luminal breast cancers.

Cost-effective management of gynecomastia

BACKGROUND Routine endocrine screening of idiopathic gynecomastia has been advocated, but may not be cost effective. We carried out a cost-benefit analysis of this approach. METHODS A retrospective study (1992 to 1997) of 87 adult males with symptomatic gynecomastia was performed. RESULTS Thirty-four (39%) patients had extrinsic causes; 53 (61%) were considered idiopathic. Forty-five idiopathic cases underwent endocrine testing: beta human chorionic gonadotropin alone, 16; and beta human chorionic gonadotropin, LH, estradiol, testosterone+/-testicular ultrasound, 29. One (2%) occult Leydig cell testicular tumor was detected. Forty-four patients had normal studies and remain well after local excision. CONCLUSION Routine endocrine evaluation of idiopathic gynecomastia is rarely productive; such testing is best done selectively.

Hepatic complications of breast cancer

Hepatic disease associated with breast cancer is common and can result from metastatic spread of the tumour to the liver, or can be caused by systemic treatment with chemotherapeutic or antiendocrine agents. Metastatic disease to the liver can present clinically and pathologically in various ways. Little is known as to why breast cancer can sometimes present as liver dominant disease or with liver involvement as a late event in the disease course. However, there are many postulations involving metastasis organotropism, which might offer future insight. The mainstay of treatment for hepatic metastases continues to be systemic therapy, but several locoregional adjunct therapies exist. Despite these therapies, liver metastasis from breast cancer is associated with a poor prognosis. Ongoing research of the mechanisms and tropism of liver metastasis from breast cancer will hopefully result in improved targeted therapies to reduce their incidence and improve outcomes when they arise.

Enhanced insulin signaling via Shc in human breast cancer

Insulin is a mild mitogen and has been shown to potentiate mitogenic influence of other growth factors. Because hyperinsulinemia and/or overexpression of insulin receptors have been linked to development, progression, and outcome of breast cancer, we attempted to evaluate the mechanism of these associations. We have compared the expression of insulin receptors and the magnitude of insulin signaling in breast tumors and adjacent normal mammary tissue samples obtained from 20 patients. We observed that insulin binding more than doubled in the tumors as compared with the normal tissue (P <.01 by paired t test). Insulin signaling to Shc, judged by the magnitude of its phosphorylation, was also significantly enhanced in the tumors. In contrast, the phosphorylation of the insulin-receptor substrate-1 (IRS-1), Akt, and mitogen-activated protein (MAP) kinase were identical in the tumorous and normal mammary tissues. Finally, tumors displayed significantly increased amounts of farnesylated p21 Ras and geranylgeranylated Rho-A (P <.01), consistent with Shc-dependent activation of farnesyl (FTase) and geranylgeranyl transferases (GGTase) in the tumor tissue. We conclude that the mechanism of the mitogenic influence of insulin in breast cancer may include increased expression of insulin receptors, preferential hyperphosphorylation of Shc, and increased amounts of prenylated p21 Ras and Rho-A in tumor tissue as compared with adjacent normal mammary tissue.

Discovery and verification of protein differences between Er positive/Her2/neu negative breast tumor tissue and matched adjacent normal breast tissue

This study was designed to quantify and identify differences in protein levels between tumor and adjacent normal breast tissue from the same breast in 18 women with stage I/II ER positive/Her2/neu negative invasive breast cancer. Eighteen separate difference gel electrophoresis (DIGE) gels were run (1 gel per patient). Relative quantification was based on DIGE analysis. After excision and tryptic digestion, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and peptide mass mapping were used to identify protein spots. Two hundred and forty-three spots were differentially abundant between normal and cancer tissues. Fifty spots were identified: 41 were over abundant and nine were less abundant in cancers than in normal breast tissue. Western blotting provided independent confirmation for three of the most biologically and statistically interesting proteins. All 18 gels were replicated by another technician and 32% of the differentially abundant proteins were verified by the duplicate analysis. Follow-up studies are now examining these proteins as biomarkers in blood.

Sentinel node biopsy: ALARA and other considerations.

For a majority of solid tumors, the most powerful and predictive prognostic factor is the status of the regional lymph nodes. Sentinel lymph node sampling continues to gain in popularity as patients and their physicians seek to avoid the potential morbidity associated with standard axillary node dissection. Lymphoscintigraphy, one of the recently explored techniques of lymphatic mapping, involves pre-operative intradermal or subcutaneous administration of a radiopharmaceutical. While this approach is gaining widely spread acceptance, there is still a lack of consensus on which radiopharmaceutical agent has the most ideal properties. By far, the most commonly used agents are 99mTc labeled colloids, but other agents are also used clinically and are under investigation or development worldwide. A number of other clinical, technical, dosimetric, and logistical considerations regarding this procedure remain. They include questions such as who should be performing the procedure, what precautions to take during surgery, how to better isolate “hot” nodes and thus improve the efficacy of determining metastases to the draining lymph node, what precautions to take when handling surgical specimens, etc. There is clearly a need to review as low as reasonably achievable considerations and other issues that arise as this technique evolves and finds its role in the evaluation of various types of cancers. This paper, based on our own experiences and those of others, fills this gap.

Unique Profile of Adenoid Cystic Carcinoma: A Triple Negative Breast Tumor With Paradoxical Features, a Case Report and Review of Literature

Adenoid cystic carcinomas (ACC) of the breast are uncommon tumors that are not well characterized and have seemingly paradoxical features. We report the case of a deceptively well circumscribed adenoid cystic carcinoma with multiple additional foci of invasive tumor. Immunohistochemical studies revealed an estrogen receptor (ER), progesterone receptor (PR), Human epidermal growth factor Receptor 2 (HER2/ neu ) negative tumor (triple negative tumor) with immunoreactivity for p63 and, CD 117 (c-kit). p53 was not over expressed. Gene studies did not reveal mutations in the commonly mutated exons of the TP53 and KIT genes. The unique and paradoxical features of this tumor are presented along with a review of the literature. * ACCs : adenoid cystic carcinomas ER : estrogen receptor PR : progesterone receptor HER2/ neu : Human epidermal growth factor Receptor 2 IHC : immunohistochemistry PCR : polymerase chain reaction TNBCs : triple negative breast carcinomas MGA : microglandular adenosis

Immunomodulatory therapy in multiple sclerosis and breast cancer risk: a case report and literature review.

Multiple sclerosis (MS) is an autoimmune inflammatory disease of the central nervous system frequently complicated by devastating neurologic symptoms and progressive disability. Much progress has been made in the development of immunomodulating drugs to help fight the progression of MS. These drugs are believed to work by interacting with various immune system components to reduce the amount of autoimmune destruction to the nervous system. We report 2 cases of women with MS on immunomodulatory therapy who presented with locally advanced breast cancer with aggressive biologic phenotypes and exceptionally poor outcomes. We consider the potential for an increased risk of developing a poorer-prognosis breast cancer as a result of concomitant immunomodulatory effects of the previous MS treatment, particularly the effects the drugs are reported to have on regulatory T cells, and therefore present these cases and a review of the current literature. Current data in the literature reflect the need for further study in ascertaining the risk of biologically poor-prognosis breast cancer development in patients with MS treated with immunomodulatory therapy.

Abstract A98: Young women's breast cancer demonstrates increased immune suppression through ciculating regulatory T cells and myeloid-derived supressor cells independent of stage or subtype.

Background: Women age 20-40 have a higher ten year risk for developing breast cancer than the five other leading cancers in this gender and age group combined. Women currently in their 30s have a 1/203 risk for breast cancer in the ensuing ten years. Cancers diagnosed in this age population of breast cancer have a higher risk for death for reasons that are not fully identified. Regulatory T cell (Tregs) and myeloid derived suppressor cells (MDSC) have a role in suppressing anti-tumor immunity. MDSC function through Jak/Stat pathway activation by generation of reactive oxygen species and/or arginase-1, and have been correlated pre-clinically with cancer progression and poorer prognostic features and outcomes. Our Young Womens Breast Cancer Translational Program seeks to identify immunologic differences potentially contributing to the poorer prognosis and potential targets for development of immunomodulatory treatment. Hypothesis: We hypothesized that newly diagnosed, treatment-naive young breast cancer cases would have higher percentage of circulating Tregs and MDSC with T cell suppressive function that similar age women without breast cancer. Methods: We conducted an IRB-approved, prospective translational study of women age 40 and under both affected and unaffected by breast cancer. Exclusion criteria included pregnancy, known autoimmunity or immunosuppressive medications, or other cancers. Tregs and MDSC were isolated from peripheral blood and phenotyped through standard protocols. The ability of MDSC to suppress T cell function were tested in co-culture assays for expression of activation marker CD25, production of gamma-interferon and production of arginase-1 and generation of reactive oxygen species. Results: As expected, Tregs were in higher number in young women with breast cancer than unaffecteds. Contrary to prior publications no difference was detected in number of MDSC identified in breast cancer (n=61) versus unaffected subjects (n=18). However, statistically significant increase in the MDSC ability to suppress T cell activation was identified in the breast cancer cohort with decreased CD25 expression and gamma-interferon production. MDSC from young women with breast cancer also secreted higher arginase-1 but not ROS from their MDSC. Level of MDSC suppression was independent of stage or biologic subtype of the breast cancer and did not correlate with the level of Tregs in the patient. Conclusions: The presence of equal rather than higher numbers of MDSC circulating in young onset breast cancer versus unaffected subjects was unexpected in comparison to prior publications. Our results may differ due to our larger sample size or the alignment by gender and age between the cohorts. Also, these are the first data on Tregs and MDSC to specifically focus on a young female population. Despite the equal numbers, MDSC from the breast cancer cohort demonstrated increased ability to suppress T cell function which was independent of cancer stage, biologic subtype or number of Tregs in the subject. These data suggest a functional difference to MDSC in the young breast cancer-bearing woman that could be exploited in even early stage breast cancer and that is not directly dependent on Tregs numbers. Moreover, the data indicate that unaffected young women have a higher than expected number of MDSC available for cancer to usurp and induce immune suppression. Identification of secretion of arginase-1 as a potential mechanism of immune suppression in young womens breast cancer warrants further investigation. Citation Format: Virginia F. Borges, Oscar Ramirez, Michelle Borakove, Elizabeth Manthey, Christina Finlayson, Anthony Elias, Martin McCarter, Kimberley Jordan, Jennifer Diamond, Nicole Kounalakis. Young womens breast cancer demonstrates increased immune suppression through ciculating regulatory T cells and myeloid-derived supressor cells independent of stage or subtype. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology: Multidisciplinary Science Driving Basic and Clinical Advances; Dec 2-5, 2012; Miami, FL. Philadelphia (PA): AACR; Cancer Res 2013;73(1 Suppl):Abstract nr A98.

Abstract P4-04-06: Young women's breast cancer is characterized by increased immune suppression through circulating myeloid derived supressor cells

Background: Women age 20–40 have a higher ten-year risk for developing breast cancer than the five other leading cancers in this gender and age group combined. Women currently in their 30s have a 1/203 chance of developing breast cancer over the next ten years. Moreover, cancers diagnosed in younger women have a signficantly higher risk of death for reasons that are not fully identified. Myeloid derived suppressor cells (MDSC) have a role in suppressing anti-tumor immunity through Jak/Stat pathway activation by generation of reactive oxygen species (ROS) and arginase-1, and correlate preclinically with cancer progression and in human canceres with poorer prognositic features and outcomes. Our Young Women9s Breast Cancer Translational Program seeks to identify immunologic differences potentially contributing to the poorer prognosis and potential targets for development of immunomodulatory treatment. Hypothesis: We hypothesized that newly diagnosed, treatment-naive young breast cancer cases would have higher percentage of circulating MDSC with T cell suppressive function than similar age women without breast cancer. Methods: We conducted IRB approved, prospective translational studies of women age 40 and under both affected and unaffected by breast cancer. Exclusion criteria included pregnancy, known autoimmunity or immunosuppressive medications, or other cancers. MDSC were isolated from peripheral blood and phenotyped through standard protocols. T cell suppressive abilites were tested in coculture assays for expression of activation markers CD25 and CD69, and production of gamma-interferon. Identification of mechanism of suppression was assayed through secretion of arginase-1and generation of ROS. Results: No difference in percentage of MDSC was identified between young women with breast cancer (n = 61) and unaffected subjects (n = 18). However, a statistically significant increase in the MDSC ability to suppress T cell activation was identified in the breast cancer cohort with diminished CD25 and CD69 expression and diminished production of gamma-interferon. MDSC from young breast cancer subjects secreted significantly more arginase-1 from MDSC. No significant difference in generation of ROS was found between the two cohorts. Conclusions: The presence of equal rather than higher numbers of circulating MDSCs between the young breast cancer versus unaffected subjects was unexpected in comparison to prior publications on MDSC in cancer. Our results may differ due to our larger sample size or the alignment by gender and age the cohorts being more representative. Also, these are the first data on MDSC in a young female population. Despite equal numbers of MDSCs, the functional analyses demonstrate MDSC isolated from breast cancer have enhanced T cell suppression capabilities compared with normal controls. These data provacatively suggest a functional difference inherent to MDSC in the young breast cancer-bearing host. Moreover, they indicate that normal young women have realtively high levels of MDSC that are available for cancer to ursurp and induce immune suppression. Identification of secretion of arginine-1 as a potential mechanism of immune supporession in young women9s breast cancer warrants further investigation. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-04-06.