bluuebunny/crossref_metadata_2025 huggingface dataset:
Clinicopathological characteristics of subsequent breast cancers in patients with benign breast disease
Authors: Santana-Davila R. , Visscher D. W. , Vachon C. M. , Frost M. , Vierkant R. A. , Anderson S. S. , Degnim A. C. , Hartmann L. C.
Published: August 2020

1061

Background: Benign breast disease (BBD) is a significant risk factor for breast cancer (BC); however little is known about the aggressiveness of the BCs these patients develop. Shared features between select atypias and low-grade DCIS have led some to speculate that a history of BBD portends the development of well-differentiated BCs. Methods: The Mayo BBD cohort includes 9,376 women who had benign breast biopsies from 1967–1991. Data on BC events were obtained from medical records and questionnaires. Tissue from the BCs was reviewed by a single breast pathologist (DWV). Results: Our cohort included 6,244 patients with nonproliferative disease (NP), 2,801 women with proliferative disease without atypia (PDWA), and 331 with atypical hyperplasias (AH). With a median of 18 years of follow-up, 799 patients with BBD have developed breast cancer, 416 initially had NP, 313 had PDWA, and 70 AH. BC tissue was available for 703 of the women. The cancers were invasive ductal in 76.1% (n = 535), invasive lobular in 9.4% (n = 66), and DCIS alone in 14.5% (n = 102) patients. Grade is currently available for 500 women, and is well differentiated in 29.4% (n = 150), moderately differentiated in 42.7% (n = 218) and poorly differentiated in 27.8% (n = 142). In 537 cases the malignant tissue had a concurrent benign component consisting of NP in 24% (n = 129), PDWA in 25.1% (n = 135), and AH in 50.8% (n = 273). In regards to tumor size, 69.6% had T1 tumors, 24.9% T2 tumors, and 5.5% T3 disease. Metastasis to lymph nodes occurred in 25% of patients. Median time to BC was 12.3 years and did not differ across the different benign entities. Breast cancer developed within 5 years from the initial BBD in 163 women (20%). In this group, tumor size was greater (p = 0.02), and there were more poorly differentiated tumors (p = 0.006) than in those who later developed BC. Conclusions: In the BCs that have developed in our BBD cohort, we do not see a preponderance of well-differentiated, good-risk BCs. Patients diagnosed within 5 years of BBD appear to have BCs with more aggressive features.

No significant financial relationships to disclose.
First-line lapatinib combined with letrozole versus letrozole alone for hormone receptor positive (HR+) metastatic breast cancer (MBC): Subgroup analyses of borderline FISH+, IHC 2+, HER2 unknown (UNK), and treatment-naive (TN) populations from EGF30008
Authors: O'Rourke L. , Pegram M. , Press M. , Pippen J. , Pivot X. , Gomez H. , Florance A. , Maltzman J. , Johnston S. , Johnston S.
Published: August 2020

1062

Background: This double-blind, placebo-controlled, phase III trial assessed the benefit of adding lapatinib, an oral EGFR/HER2 tyrosine-kinase inhibitor, to letrozole alone in patients (pts) with HR+ MBC. The previously reported primary endpoint, investigator (INV) assessed PFS in HER2+ tumors, showed a significant benefit from dual therapy. Median PFS in the HR+ HER2+ population increased from 3 months (mo) in the letrozole/placebo group to 8.2 mo in the letrozole/lapatinib group [Hazard Ratio (95% CI)=0.71 (0.53,0.96), stratified log rank p = 0.019]. The HER-2-ve population did not derive benefit from the combination. Benefit from combined treatment was evaluated in a number of pre-planned exploratory subsets, including a noted trend in the HER2-ve population who progressed within 6 mo of receiving prior tamoxifen. Methods: 1286 pts were randomized to letrozole/lapatinib or letrozole/placebo. HER2 positivity was defined by a positive FISH ratio or by immunohistochemistry (IHC) 3+ in a central laboratory. INV assessed PFS in the sub-populations were analyzed using Kaplan-Meier with stratified log rank to compare treatment arms. These included tumor samples that were FISH borderline1.8–2.2 (n = 52), IHC 2+ (n = 215), HER2 status UNK (n = 115), and neo/adjuvant TN (n = 656). Results: INV assessed PFS demonstrated no significant prolongation for dual therapy for any of the exploratory populations [Hazard Ratio: (95%CI), p-value]; FISH 1.8–2.2 [1.03 (0.55, 1.95), p = 0.918]; IHC 2+ [1.13 (0.82, 1.57), p = 0.441]; HER2 UNK [0.71 (0.45,1.11), p = 0.126]; TN [0.88 (0.73, 1.07), [p = 0.199]. Conclusions: The combination of letrozole and lapatinib did not significantly improve PFS in any of the pts with lower levels of HER2 expression (borderline FISH, IHC 2+, or HER2 UNK) or in TN pts. These data confirm the HER2-ve result previously reported and substantiate that only tumors with the target benefit from the addition of a targeted therapy.

[Table: see text]
A phase I/II study of CR011-vcMMAE, an antibody-drug conjugate, in patients (pts) with locally advanced or metastatic breast cancer (MBC)
Authors: Peacock N. , Saleh M. , Bendell J. , Rose A. A. , Dong Z. , Siegel P. M. , Crowley E. , Simantov R. , Vahdat L.
Published: August 2020

1067

Background: Glycoprotein NMB (GPNMB), also known as osteoactivin, has been shown to regulate metastasis of breast cancer in vivo. CR011-vcMMAE, a fully-human monoclonal anti-GPNMB antibody conjugated to the tubulin inhibitor monomethylauristatin E, is safe and active in pts with advanced melanoma. This is the first study of CR011-vcMMAE in breast cancer. Methods: Eligible pts with MBC had ≥ 2 prior chemotherapy regimens, including a taxane, an anthracycline, and capecitabine; and ECOG PS ≤ 2. Doses were escalated to 1.88 mg/kg IV q3w (the maximum tolerated dose [MTD] in melanoma) using a standard 3+3 design. Immunohistochemistry (IHC) with goat polyclonal antibody to GPNMB was performed on pt biopsy specimens and on tissue microarrays containing normal breast, DCIS, breast tumor and lymph node metastases. Results: 10 pts with MBC (median age 57, range 36 - 69) had a median of 7 prior regimens and were treated with CR011-vcMMAE for a median of 2 cycles (range 1–4). In the first 2 pts at 1.34 mg/kg, dose limiting toxicity of worsening peripheral sensory neuropathy was observed. Pts with baseline neuropathy worse than grade 1 were subsequently excluded. Pts were treated at 1.0 mg/kg (n = 3), 1.34 mg/kg (n = 5), and 1.88 mg/kg (n = 2); enrollment at 1.88 mg/kg is continuing. Other adverse events (AEs) were grade 1/2 anorexia and pain in 4 pts; diarrhea, rash, fatigue, and neuropathy in 3 pts; and grade 3 rash in 1 pt. Evidence of antitumor activity has been observed. A response of 37% tumor shrinkage was seen in a pt after only 2 cycles and is ongoing. A second pt had a 51% reduction after 2 cycles, but had PD after 12 weeks. Breast tumor samples were more likely to stain positive for GPNMB than normal breast tissues. Conclusions: CR011-vcMMAE 1.34 mg/kg IV q3w is well-tolerated in heavily pretreated pts with MBC. The 1.88 mg/kg q3w dose is being assessed and phase II expansion is planned at the MTD. IHC of pt tumor specimens is being evaluated.

[Table: see text]
Effect of race on the safety and efficacy outcomes of gemcitabine plus paclitaxel treated patients with metastatic breast cancer (MBC): Analyses from a phase III trial
Authors: Wang Y. , Tai D. , Zhao L. , Gill J. , Obasaju C. K.
Published: August 2020

1070

Background: Population-based studies often attribute racial disparities in breast cancer outcomes to differences in access to treatment, quality of care, or other socioeconomic factors. In a controlled clinical trial setting, these systemic differences between races should be minimal. To evaluate the potential impact of race on outcomes in a controlled clinical setting, we retrospectively analyzed data from a phase III trial (B9E-MC-JHQG; NCT00006459 ) of patients (pts) with MBC. Methods: Analyses were performed on the JHQG trial database after 440 total pt deaths for both study arms. Demographics, safety, and efficacy were analyzed using safety population data from 3 racial groups: Caucasian (CA), Asian (AS), and Hispanic (HP). CA was the reference for all pair-wise comparisons. The logistic model was used to calculate odds ratios for tumor response and the Cox model was used to calculate hazard ratios for time-to-event parameters, adjusting for significant prognostic factors. Results: We report analyses of the gemcitabine (G) + paclitaxel (T) treatment arm. Demographics were balanced across the 3 groups with the exception that ER+/PR+ status was lower for AS compared to CA and HP; unknown ER/PR status was higher for AS. AS had significantly less neutropenia, fatigue, and nausea, but more anemia compared to CA and HP. Median number of treatment cycles completed was lower, but mean dose intensities for G and T were slightly higher, for AS. Response rate and progression-free survival were similar in the 3 groups. Overall survival (OS) and post-study chemotherapy (PSC) were significantly reduced for AS. Conclusions: Our analysis suggests that AS pts were better able to tolerate GT therapy compared to CA and HP pts. However, AS pts had the poorest OS outcome of the 3 racial groups, potentially due to reduced participation in PSC.

[Table: see text]

[Table: see text]
Imatinib-induced changes in gene expression and the metastatic potential of human breast carcinoma cells
Authors: Lorico A. , Anzanello F. , Rappa G.
Published: August 2020

1074

Background: Imatinib mesylate (imatinib) is a potent and selective inhibitor of the tyrosine kinases, Bcr-Abl, c-Kit and platelet-derived growth factor receptors (PDGFRs). Since its advent for the successful treatment of chronic myelogenous leukemia in 2001, the clinical efficacy of imatinib has been investigated in many other human malignancies, including breast cancer. Based on recent reports that chemotherapy selects more invasive and metastasizing cells, we have hypothesized that exposure of breast cancer cells to imatinib could enhance their malignant behavior. Methods: MA-11 breast carcinoma cells, originating from bone marrow micrometastases, were exposed to imatinib in vitro for seven days. After four days of recovery in drug-free medium, biological properties and gene expression pattern were compared with those of the parental cell line. In a separate set of experiments, the effects of in vivo administration of imatinib to athymic nude (nu/nu) mice carrying MA-11 tumors were investigated. Results: In vitro, imatinib treatment increased the motility and invasiveness of the breast cancer cells, and induced over-expression of drug transporters and of a set of genes associated with aggressive and metastatic behavior (Table). In vivo, nu/nu mice subcutaneously implanted with MA-11 cells and treated with nine daily intraperitoneal doses of 60 mg/Kg imatinib developed with greater frequency distant organ metastases vs. control mice implanted with MA-11 and treated with the vehicle alone. Conclusions: Our data caution against the clinical use of imatinib in breast cancer; imatinib-selected breast cancer cells represent an important tool to investigate the pro-metastatic role of differentially expressed genes.

[Table: see text]

No significant financial relationships to disclose.
Phase II trial of the histone deacetylase inhibitor, vorinostat, to restore hormone sensitivity to the antiestrogen tamoxifen in patients with advanced breast cancer who progressed on prior hormone therapy
Authors: Munster P. N. , Lacevic M. , Thomas S. , Christian C. , Ismail-Khan R. , Melisko M. , Rugo H. , Minton S. E.
Published: August 2020

1075

Background: Modulation of estrogen receptor signaling is one of the most successful modalities in the treatment of estrogen receptor (ER) positive breast cancers. However, hormone therapy resistance remains a challenging problem. Histone deacetylases (HDAC) have emerged as a recent strategy to interfere with hormone receptor signaling by modulating the estrogen and progesterone receptor. Here we report the findings from a phase II trial evaluating the HDAC inhibitor, vorinostat and the anti-estrogen, tamoxifen, placed in context with correlative studies. Methods: Patients with ER-positive metastatic breast cancer who progressed on prior hormonal therapy and up to three chemotherapy regimens were treated with 400 mg vorinostat daily for three of four weeks and 20 mg tamoxifen daily, continuously. Histone acetylation was evaluated in peripheral blood mononuclear cells. Results: To date, 29 patients [median age 53 years (34–71)] have been treated with the combination. Pulmonary emboli were observed in 2/29 (7%) patients. Other grade 3/4 toxicities included fatigue (5/29, 17%), anorexia (2/29, 7%), diarrhea (1/29, 3%), nausea/vomiting (3/29, 10%), hypokalemia (1/29, 3%), liver enzyme elevation (1/29, 3%), lymphopenia (5/29, 17%) and leukopenia (4/29, 13%). Predominant grade 2 toxicities included fatigue, nausea/vomiting/diarrhea, hyperglycemia, anorexia, and myelosuppression. Six (21%) patients had an objective response, and 3/29 (10%) additional patients had disease stabilization for >6 months; 13/29 (45%) patients had at least one prior chemotherapy regimen for metastatic disease, 28/29 (92%) patients had progressed after exposure to at least one and 16/29 (55%) patients after two aromatase inhibitors, and 18/29 (62%) patients had received prior adjuvant tamoxifen. H3 and H4 histone acetylation was seen at day 8 suggesting adequate vorinostat plasma levels. Histone acetylation and baseline HDAC expression will be presented. Conclusions: These findings suggest that the addition of an HDAC inhibitor to tamoxifen in patients who have progressed on prior hormonal therapy and/or adjuvant tamoxifen may restore hormone sensitivity.

No significant financial relationships to disclose.
Evaluation of Mena isoforms as a surrogate for epithelial mesenchymal transformation and erlotinib resistance in breast carcinoma
Authors: Ghiuzeli C. M. , Roussos E. T. , Wyckoff J. B. , Sun D. , Wang Y. , Patsialou T. , Goswami S. , Gertler F. B. , Condeelis J. S.
Published: August 2020

1078

Background: Epithelial to mesenchymal transition (EMT) is an important step in invasiveness and has been shown to correlate with metastatic potential in several cancer cell lines, including breast carcinoma. However, given the heterogeneity of tumors in vivo, EMT has not been a reliable marker of metastatic potential in cancer patients. Mena, a member of the enabled (ena)/vasodilator-stimulated phophoprotein (VASP) family, which controls cell motility, is upregulated in the invasive subpopulation of breast cancer cells. Mena is alternatively spliced to include one of four exons: +, ++, invasive (INV), or 11a. In the presence of MenaINV, tumor cells are able to invade even at epidermal growth factor (EGF) concentrations that would otherwise be undetectable by the tumor cells. Currently, there are several clinical and genetic characteristics which can predict sensitivity to erlotinib, an EGF receptor inhibitor, but further studies are necessary. Methods: The animal models used were the polyoma middle T antigen (PyMT) transgenic mouse and severe combined immunodeficiency (SCID) xenografted tumors derived from injection with a human breast carcinoma line, MDA-MB-231, and a rat adenocarcinoma line, MTLn3, with forced expression of MenaINV and Mena11a. Invasive cells and average primary tumor cells were collected using the in vivo invasion assay and Fluorescence activated cell sorting (FACS), respectively, and imaged via immunofluorescence. Results: We show that in both the PyMT and MDA-MB-231 animal models, invasive cells are significantly more mesenchymal than average primary tumor cells, indicating they have undergone EMT. The MTLn3-MenaINV cells are more mesenchymal-like, whereas the Mena11a are more epithelial-like. We also expect that cells with a high Mena INV/11a ratio invade despite the presence of erlotinib.

No significant financial relationships to disclose.
Effect of dasatinib on the activity of trastuzumab in HER2-overexpressing breast cancer cells
Authors: Seoane S. , Montero J. C. , Pandiella A. , Ocana A.
Published: August 2020

1084

Background: Breast cancer is the most common neoplasia in women. Although important advances in its treatment have been made along the last years, in the metastatic setting this disease remains incurable. One group of molecules involved in breast cancer genesis/progression are tyrosine kinases. HER-2 is a receptor tyrosine kinase that is overexpressed in a subset of patients with breast cancer. Targeting of HER-2 with trastuzumab a monoclonal antibody that acts on HER-2 has demonstrated clinical benefit. However, as a single agent its antitumoral activity is limited. Methods: Here we have investigated whether the action of trastuzumab, can be potentiated by dasatinib, a small molecule inhibitor that acts on several receptor and non-receptor tyrosine kinases. To this end, we have used cancer cell lines and in vivo xenografted mouse models. MTT proliferation and other apoptosis assays, biochemical and microarray experiments were performed to investigate the molecular bases of the action of the combined trastuzumab plus dasatinib treatment. Results: Combination of trastuzumab and dasatinib was synergic on breast cancer cells overexpressing HER-2, but not on cells with a normal complement of its receptor. In vivo xenograft studies confirmed the antitumoral action of this drug combination. Studies on the mechanism of action showed that this drug combination triggered apoptosis in HER-2 overexpressing cells. Apoptosis was evidenced by increased Annexin V staining, and was found to occur in a caspase-independent manner. In addition, an action of the drug combination on cell cycle could be identified, and the drugs decreased cyclins E1/2, B, and Cdk2, in addition to increasing p27. Microarray as well as biochemical analyses showed that the drug combination provoked a decrease in HER receptor phosphorylation, followed by interruption of downstream signalling. These changes resulted in alterations in the levels of proteins involved in DNA repair. Moreover, treatment with the drug combination caused a DNA damage response as evidenced by increased γH2AX. Conclusions: The effective antitumoral action of the drug combination opens new avenues for the clinical exploration of this drug combination in breast cancer patients overexpressing HER-2.

No significant financial relationships to disclose.
Preliminary results of a randomized phase II study of paclitaxel and bevacizumab ± gemcitabine as first-line treatment for metastatic breast cancer
Authors: Hoelzer K. L. , Brufsky A. , Hainsworth J. , Beck J. T. , Whorf R. , Keaton M. , Kroener J. , Krill-Jackson E. , Hu S. , Bromund J.
Published: August 2020

1089

Background: The addition of bevacizumab (B) to paclitaxel (P) results in a significant improvement in PFS in pts with metastatic breast cancer (MBC) (Miller K, et al. New Engl J Med 2007). A randomized Phase II trial examining the efficacy and safety of adding gemcitabine (G) to the PB doublet has completed enrollment. Reported here are preliminary efficacy and safety results. Methods: This is a US, multicenter, randomized, superiority trial. Eligible pts have locally advanced or metastatic breast cancer, ECOG PS 0 or 1, and no prior cytotoxic therapy for metastatic disease. Prior adjuvant or neoadjuvant treatment with a taxane or endocrine therapy is allowed. Pts are randomized to receive P 90 mg/m2 on Days 1, 8, and 15, followed by B 10 mg/kg on Days 1 and 15 of a 28-day cycle, or the same regimen plus G 1,500 mg/m2 on Days 1 and 15. Primary endpoint is response rate according to RECIST criteria. Results: Between May 2006 and February 2008, 189 women were randomized to treatment. The table below summarizes currently available results. Grades 1–2 alopecia occurred in 28% of pts in the PB arm and in 38% of pts in the PB+G arm. One pt (2%) in the PB arm experienced a Grade 3 nosebleed. Grades 3 and 4 thrombotic events occurred respectively in 0% and 2% of pts in the PB arm, and in 3% and 2% of pts in the PB+G arm. Four pts (7%) in the PB arm and 3 pts (5%) in the PB+G arm discontinued due to treatment-related AEs. Three on-study deaths have occurred, none deemed related to study treatment. Conclusions: Study follow-up is ongoing. Full results will be available at the time of the meeting. Therapy with PB ± G is feasible and does not appear to be associated with significant bleeding or thrombotic events. As expected, the addition of G to the PB doublet appears to increase the incidence of neutropenia in pts with MBC.

[Table: see text]

[Table: see text]
Efficacy results from a multicenter phase II noncomparative two-arm pilot trial of bevacizumab with anastrozole or fulvestrant as first-line endocrine therapy for metastatic breast cancer
Authors: Rubin M. S. , Barton J. , Shipley D. , Arrowsmith E. , Peacock N. , Hart L. , Evans J. , Vasquez E. , Burris H. A. , Yardley D. A.
Published: August 2020

1091

Background: Estrogen modulates angiogenesis via effects on endothelial cells with subsequent induction of vascular endothelial growth factor (VEGF). VEGF promotes tumor growth and is associated with poor response to antiestrogen therapy. This trial was designed to evaluate the progression-free survival (PFS) of bevacizumab (B) in combination with anastrozole (A) or fulvestrant (F) as first-line endocrine therapy (ET) in metastatic breast cancer (MBC). Methods: Eligibility criteria: no prior hormonal or chemotherapy for MBC, measurable or evaluable disease, normal LVEF, post-menopausal. Treatment: Arm A: anastrozole 1 mg po QD in pts who were a) ET naïve, b) ≥ 12 months from adjuvant ET, and c) intolerant of or progressed on prior tamoxifen. Arm B: fulvestrant 500 mg D1 and 250 mg D15 IM loading dose followed by 250 mg q28 days in pts who were a) < 12 months from adjuvant aromatase inhibitors (AIs), b) intolerant of or progressed on AIs, and c) MD's discretion. Bevacizumab 10 mg/kg IV D1 q2 weeks was given in both arms. Trastuzumab permitted in HER-2+ pts only. Response assessments were q8 weeks; pts were treated until disease progression or toxicity. Results: 79 pts were enrolled fromNovember 2006 to November 2008. 42 pts are evaluable for response and toxicity, Arm A - 25 pts and Arm B - 17 pts. Median age was 64, ECOG PS 0 - 55%, 1- 43 %, adjuvant chemo 27%, adjuvant hormonal -38%, hormone receptor status: ER+/PR+ 80%, ER+/PR- 14%, ER-/PR+ 2 %. HER-2+ 5 pts, 31% had ≥ 2 metastatic disease sites predominately lung and bone only disease - 40%. Median # cycles - 4. 24% achieved a partial response and 57% stable disease; 7 pts progressed. G3 hypertension (12%) was the most common toxicity. Median PFS for Arm A was 16.3 months and has not yet been reached for Arm B. Conclusions: Bevacizumab in combination with anastrozole or fulvestrant is feasible and well tolerated with no unanticipated toxicities. The addition of bevacizumab resulted in prolongation of the median PFS to16.3 months with anastrozole as compared to the 7–9 month historical control PFS reported for first-line AI monotherapy in MBC. Further evaluation of bevacizumab endocrine combinations is warranted.

[Table: see text]
A phase II trial of gemcitabine combined with vinorelbine as first-line chemotherapy for metastatic breast cancer
Authors: Meshref M. M. , Samir S. , Saad E. , Gouda Y. , Elmesidy S. , Koheil H. , Elzawahri H. , Kandil A. , Abdelhamid T. , Zaki M.
Published: August 2020

1098

Background: MBC (metastatic breast cancer) is among the leading causes of cancer deaths in women worldwide and in Egypt. Gemcitabine has shown synergy with vinorelbine in preclinical models, and has a toxicity profile that is different from vinorelbine. Methods: This was an open-label, single-arm, non-randomised phase II study in patients with locally advanced or MBC who have been previously treated with one anthracycline with/without taxane, based regimen in the adjuvant/neoadjuvant setting. The primary objective of this study was to assess the efficacy of this combination by determining overall response rate. Secondary endpoints were the assessment of the toxicity of gemcitabine in combination with vinorelbine, as well as the time to disease progression, one-year survival. Results: Of the 74 patients enrolled, 72 patients were evaluable for primary treatment outcome (tumor response rates). Overall response rate was 42%. Four patients (6%) had complete response and 26 patients (36%) had partial response. Nineteen patients (26%) had stable disease. Among the 30 patients who had a response to treatment during the study period, the median time to disease progression was 37 weeks (range 1 -60). Median duration of response was 43 weeks (95% CI 31- unestimated). The probability of surviving one year from the Kaplan-Meier estimate for 12 months is 77% (95% CI 64%-86%). Mean percentage of dose intensity of gemcitabine and vinorelbine received was 86% (SD 13%). Mean number of cycles received was 4.4 (SD 1.8). Grade III-IV neutropenia was reported to be 10%, thrombocytopenia was 1.4%, and febrile neutropenia (with documented infection) was 1.4%. As for the clinical toxicities, the most common grade III-IV toxicities were nausea (24%) and diarrhea and stomatitis (11%). Serious clinical adverse events; 5 (7%) hospitalizations for adverse events mainly due to anemia, febrile neutropenia, septic shock and hepatic failure. There were 2 platelet transfusions and 14 RBC transfusions. Conclusions: With a disease control rate of 68%, gemcitabine/vinorelbine demonstrated to have a promising efficacy and at the same time good tolerability in metastatic breast cancer patients.

[Table: see text]
Use of a murine model of NSCLC to evaluate the role of the microRNA-200 family in regulating EMT and metastasis
Authors: Gibbons D. L. , Lin W. , Creighton C. , Zhang S. , Lozano G. , Kurie J.
Published: August 2020

11006

Background: Non-small cell lung cancer (NSCLC) is the leading cause of cancer death worldwide, primarily due to metastatic disease. Unfortunately, we lack a clear understanding of the molecular and cellular basis for lung cancer metastasis, partly because the experimental study is hampered by lack of good models. Methods/Results: To address this deficiency we have developed an experimental murine model of metastatic NSCLC using cell lines derived from a genetic mouse model of human lung adenocarcinoma that develops metastatic disease owing to the expression of K-rasG12D and p53R172H. An expression signature derived from the spontaneous metastatic tumors in these animals is prognostic when applied to a large series of early-stage patient tumors, illustrating that the model recapitulates features of the human disease. Combined mRNA and microRNA profiling of highly metastatic subcutaneous tumors versus non-metastatic tumors revealed a signature for the epithelial-to-mesenchymal transition (including increases in known EMT-inducing transcription factors and down-regulation of genes responsible for maintenance of epithelial polarity) and associated loss of the microRNA-200 family (miR-141, 200a, 200b, 200c & 429) expression. Forced expression of the miR-200 family produced a more epithelial cell phenotype and associated changes in the ability of the cancer cells to respond to EMT-inducing stimuli such as TGFβ. We believe this to be a mechanism whereby changes in the tumor microenvironment influence tumor cell state and ability to undergo metastasis. Conclusions: New therapeutic targets to address the problem of metastatic NSCLC will only emerge from a better understanding of the tumor biology. Using this experimental animal model has revealed a role for EMT in the pathogenesis of the disease and the microRNA-200 family as regulators of the cellular switch. We are currently working to understand the microenvironment regulation of the miR-200 members. This work should provide new insights into the cell biology of tumor progression and metastasis, while generating new potential targets for therapy of metastatic disease.

[Table: see text]
Discovery of a new phospho-HER2+/FISH- molecular subtype of human breast cancer by functional pathway mapping
Authors: Wulfkuhle J. , Pierobon M. , Laird J. , Espina V. , Liotta I. L. , Esserman L. , Petricoin E.
Published: August 2020

11009

Background: Effective treatment of breast cancer through the targeting of the HER2 protein in human breast cancer represents a major advance in oncology, yet the identification of responders has relied on non-quantitative methods (IHC) that measure the erbB2 protein or indirect genomic analysis (FISH) of the erbB2 gene that cannot predict protein pathway activation status. We utilized a new quantitative protein microarray assay to measure total and phosphorylated HER2 in the context of broad-scale EGFR signal pathway mapping in order to generate a new molecular characterization scheme for human breast cancer. Methods: Pure tumor epithelium from 149 frozen pre-treatment human breast cancer tissue specimens (from the I-SPY TRIAL: CALGB 150007/150012, ACRIN 6657) were procured via Laser Capture Microdissection and protein pathway mapping was performed using Reverse Phase Protein Microarrays (RPMA) whereby the activation of 40 key signaling proteins was quantitatively measured at once. Results: While phospho-HER2 and total HER2 as measured by RPMA had excellent concordance with FISH (95%) and IHC (94%), of the 63 cases where both clinical FISH and IHC status of c-erbB2 were known, we discovered that 5/45 (11%) of the FISH-/IHC- cases had phosphorylated HER2 levels as high or higher than the FISH+/IHC positive patients. These results were confirmed and validated by independent analysis with quantitative Western Blot using a separate biopsy specimen from the same patients. Conclusions: A new molecular phenotype of human breast cancer has been identified whereby total levels of HER2 are low yet levels of the phosphorlyated receptor are very high. This molecular phenotype is not detectable by FISH analysis nor by measurement of the total HER2 protein itself. Given the central importance of phosphorylation on effective signal transduction of the EGFR family, we are planning to determine the clinical significance of this finding by retrospective analysis of banked material with outcome, and a prospective clinical trial in I-SPY 2. Support: ACRIN U01 CA079778 ; CALGB CA31964, CA33601; NCI SPORE CA58207.

[Table: see text]
Capecitabine monotherapy for patients with brain metastases from advanced breast cancer
Authors: Naskhletashvili D. R. , Gorbounova V. A. , Bychkov M. B. , Chmutin G. E. , Karakhan V. B. , Krat V. B.
Published: August 2020

1102

Background: To assess the efficacy of capecitabine monotherapy for patients with chemotherapy-pretreated advanced breast cancer with brain metastases. Methods: Patients with brain metastases from breast cancer whose disease had progressed on prior chemotherapy for advanced disease received oral capecitabine 1,000 mg/m2 bid on days 1–14 every 3 weeks until disease progression. Results: Between July 2007 and August 2008, 10 women were treated. Median age was 50 years (range 36–66 years). ECOG performance status was 1 in 7 patients and 2 in 3 patients. All had received prior chemotherapy: one line in five patients; two lines in four patients; three lines in one patient. Six patients had received prior endocrine therapy and three had received prior salvage radiotherapy to the brain. Two patients had metastases limited to the brain and the remaining eight also had extracranial metastases. All patients were assessable for response. Six achieved a partial response in the brain (CT/enhanced-contrast MRI), three showed disease stabilization, and one had disease progression as best response. Median duration of response was 4 months and median time to progression was 6 months. Seven patients are still alive and therefore median overall survival has not been reached. A total of 58 cycles of capecitabine were delivered. The most common toxicities (% of cycles) were neutropenia (G1/2 43.1%, G3/G4 8.6%), hand-foot syndrome (G1/2 25.9%, G3 8.6%), anemia (G1/2 29.3%, G3 1.7%), diarrhea (G1/2 20.6%, G3 5.2%), nausea/vomiting (G1/2 24.1%), stomatitis (G1/2 12%), thrombocytopenia (G1 8.6%), and fatigue (G1/2 6.9%). Conclusions: Our small study suggests that capecitabine has pronounced anticancer activity in patients with brain metastases from breast cancer with reasonable tolerability and easy administration as outpatient treatment. Further investigation is warranted.

No significant financial relationships to disclose.
Profiling of receptor tyrosine kinases (RTK) activation in circulating tumor cells (CTCs) in metastatic tumors using proximity mediated microarray immunoassay
Authors: Liu X. , Kim P. , Liu L. , Lee T. , Barham R. , Lin F. , Harvey J. , Ybarrondo B. , Singh S.
Published: August 2020

11024

Background: The abnormal activation of HER1 and HER2 has been linked to various types of cancer progression, and the changes in their expression status between primary tumor and CTCs have been reported to occur at a significant frequency. Methods for detecting HER1/HER2 phosphorylation in serially collected CTCs may provide valuable insight into the overall disease profile shift, and therefore lead to better selection of therapy combination for individual patient. Methods: A triple-antibody-enzyme-channeling multiplexed protein microarray platform has been developed to detect the phosphorylation on target molecules. It utilizes a unique immuno-complex formation via co-localization of two detector enzyme-conjugated-antibodies once target proteins are captured on the microarray-surface. The channeling events between two detector enzymes in proximity enabled profiling of the RTKs with a single-cell level sensitivity. The simultaneous binding of three different types of target specific antibodies for signal amplification results in extremely high assay specificity. In order to validate the method on clinical samples, CTCs from 75 cancer patients (stage 3/4) on various therapy regimens were analyzed. Results: We identified 6 patients (8%) with activated HER1, 6 patients (8%) with activated HER2 and 14 (18.5%) patients with dual RTK activation in their CTCs from various types of metastatic cancer. 25 samples from normal subjects showed no detectable HER1/HER2 activation. We also observed discrepancies between HER2 activation status in CTCs and their corresponding primary HER2-IHC status among breast cancer patients. CTCs with activated-HER2 were found in 6 out of 16 (38%) patients with HER2-negative primary breast cancer. In addition, 2 out of 5 HER2-positive breast cancer patients had CTCs with no apparent HER2-activation. Conclusions: The multiplexed-proximity mediated immunoassay successfully detected the activation of RTKs in CTCs isolated from various cancer patients. As CTCs found in metastatic stage represent the most aggressive invading cell population, serial CTC-profiling can lead to better therapy selection/adjustment and disease/treatment monitoring as there are available options to choose appropriate kinase inhibitors for RTK-targeted therapies.

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Circulating tumor cells monitored over time in lung cancer patients
Authors: Luttgen M. S. , Marrinucci D. , Lazar D. , Malchiodi M. , Clark P. , Huynh E. , Bethel K. , Bazhenova L. , Nieva J. , Kuhn P.
Published: August 2020

11025

Background: Circulating tumor cell (CTC) detection and enumeration is a valuable tool for monitoring cancer patient status and outcome. While many current techniques employ immunomagnetic-enrichment based protocols focused on the importance of a particular CTC number as the indicator of patient status or outcome, we employ a cytometric, enrichment free approach using an immunofluorescent protocol to monitor CTC counts in patients with non-small cell lung cancer (NSCLC) over the course of treatment. Methods: Eligible patients had progressive stage IV NSCLC. The histological subtypes in the 42 cases for which the data was available included adenocarcinoma (22/42), squamous cell carcinoma (6/42), large cell undifferentiated carcinoma (3/42), and non-small cell lung carcinoma not further described, poorly differentiated, or with a mixed pattern (11/42). Blood samples were collected 3 wks, 3 mo, 6 mo, 9 mo, and 1 yr after the initial sample. CTCs were identified via immunofluorescence and cytometric analysis. Patient response to therapy was determined by RECIST every 3 months between time 0 and time 12 mo. Results: 80 of 109 patient samples have CTCs (73%) and all of the 52 patients tested have CTCs. 13 of 52 patients have CTC data for time 0 and 3 wks. Only 4 of these patients (30.8%) show a correlation linking CTC count change between time 0 and 3 wks and clinical assessment. 13 patients have CTC data for time 0 and 3 mo, 10 of whom show a correlation linking CTC count change between time 0 and 3 mo and clinical assessment. 7 of the 8 patients (87.5%) showing stable or partial response at 3 mo show a decrease in CTC count between time 0 and 3 mo. Five of the 6 patients (83.3%) clinically showing progressive disease at the 3 mo time point show an increase in CTC count between time 0 and 3 mo. The patients that do not show a correlation linking CTC count change between time 0 and 3 mo and clinical assessment at 3 mo show a correlation at the 6 mo time point. Conclusions: CTCs can be effectively enumerated in metastatic NSCLC patients, with the majority demonstrating CTCs in the setting of progressive disease. The change in CTC count at 3 mo, but not at 3 wks, correlates with radiographic response to chemotherapy. Further follow-up will determine the predictive value of CTC enumeration on survival.

No significant financial relationships to disclose.
Gauging the response of circulating epithelial tumor cells (CETC) and tumor stem cell subpopulations to therapy of early-stage cancer in the individual patient
Authors: Pachmann K. , Camara O. , Runnebaum I. B. , Hoeffken K.
Published: August 2020

11026

Background: Cells released from the primary tumor persisting and recirculating in the host can lead to the formation of distant metastases. It was claimed that such cells are detectable only in a minor fraction of early-stage cancer patients but we can show that CETC are detectable and can be quantified in the peripheral blood of almost all cancer patients including early-stage solid malignancies. Methods: Using anticoagulated peripheral blood, red blood cell lysis as the only enrichment step, one centrifugation step, staining live cells with fluorochrome labelled anti-epithelial antigen as a search antibody, automated image analysis for detection of positive events and evaluation of exclusively surface located epithelial antigen on vital unfixed cells, CETC were detected in most patients with early stage cancer. Subsequently cells could be stained with anti-ALDH-antibody and in situ hybridized for HER2/neu amplification and quantified repeatedly during neo/adjuvant chemotherapy and during maintenance therapy with hormones or trastuzumab. Results: We here report the results from 497 breast cancer patients analyzed more than three times during the course of disease, 248 during neoadjuvant/adjuvant chemotherapy, 249 during trastuzumab and or hormone therapy. Different pattern of therapy response were obtained with rapidly responding CETC changes over several logs in response to chemotherapy and slow and long-lasting changes extending over several years in response to hormone therapy and trastuzumab. Stem cell like staining was seen in a minor fraction of cells (1%) in about 10% of patients. An increase in cell numbers and in the fraction of HER2/neu amplified cells was under all treatment conditions unequivocally significantly correlated to highly increased risk of relapse. Conclusions: CETC and subpopulation monitoring provides an invaluable tool for prompt gauging of systemic therapy in early stage solid tumors as a tool for therapy guidance and optimal personalized therapies to improve therapy results and spare unnecessary treatments.

No significant financial relationships to disclose.
Identification by FISH of 4 novel partner loci of PRDM16 in myeloid malignancies
Authors: Duhoux F. , Libouton J. , Bahloula K. , Ameye G. , Poirel H. A.
Published: August 2020

11037

Background: PRDM16 is a gene located on 1p36.32 that encodes for a zinc finger transcription factor and contains an N-terminal PR domain. It has been shown to be involved in the reciprocal translocation t(1;3)(p36;q21) and more rarely the t(1;21)(p36;q22) which both occur in myelodysplastic syndromes (MDS) and acute myeloid leukemias (AML). These translocations result in the overexpression of a truncated version of the PRDM16 protein that lacks the PR domain. This overexpression might play an important role in the pathogenesis of MDS and AML in blocking myeloid differentiation. This raises the question whether PRDM16 is rearranged with other partner genes and whether its sequences play the most important role in the oncogenic process attributed to those translocations. Methods: We studied 35 myeloid malignancies, 12 lymphoid malignancies and 3 undifferentiated acute leukemias with 1p36 abnormalities by fluorescent in situ hybridization (FISH) with a bacterial artificial chromosomes (BAC) contig containing 50 BAC probes on 1p36. Results: In addition to the known t(1;3)(p36;q21) (11 cases) and t(1;21)(p36;q22) (1 case) involving RPN1 andAML1/RUNX1 respectively in myeloid malignancies, we specifically found PRDM16 to be rearranged in 4 additional translocations : a t(1;12)(p36;p13) in an AML-M4, a t(1;7)(p36;p12) in a MDS, an add(1)(p36) in an AML-M2 and a t(1;2)(p36;p12) in a relapsed AML-M4. We identified the respective candidate partner loci : TEL/ETV6, IKZF1, CDH4 and a non-coding unknown sequence. Conclusions: In our series of 50 cases of hematological malignancies with 1p36 abnormalities, PRDM16 was involved in about 45% of myeloid malignancies, and was never involved in lymphoid malignancies. PRDM16 is supposed to have similar oncogenic properties as MDS1/EVI-1(3q26), another gene encoding for a zinc finger protein and acting as a transcriptional regulatory factor with 2 isoforms. Interestingly, the shortest isoform of MDS/EVI-1, lacking the PR domain, is supposed to have an oncogenic effect due to its translocation-induced upregulation in AML. Further characterization of these new partner genes and functional studies should give us more insight into the pathogenesis of AML and MDS mediated by PRDM16, and the role of its partner genes.

No significant financial relationships to disclose.
Framingham risk score after risk-reducing salpingo-oophorectomy in women at risk for hereditary breast ovarian cancer: A controlled observational study
Authors: Michelsen T. M. , Tonstad S. , Dahl A. A. , Pripp A. H. , Tropé C. G. , Dørum A.
Published: August 2020

11048

Background: Risk-reducing salpingo-oophorectomy (RRSO) effectively prevents ovarian cancer in BRCA mutation carriers and in women at risk for hereditary breast-ovarian cancer. RRSO induces immediate menopause, which may increase the risk of coronary heart disease (CHD). Our aim was to determine CHD risk using Framingham risk score and examine factors associated with this risk in women who had undergone RRSO compared to population-based controls. Methods: A sample of 326 (65% of invited) women who underwent RRSO after genetic counseling from 1980–2005 provided completed questionnaires, physical measures, and blood samples. Controls were 1,630 age-matched controls from the Norwegian Nord-Trøndelag Health Study (HUNT-2) (1995- 97). Results: Mean age in both the RRSO and control groups at survey was 54.4 years. Mean follow-up after surgery was 6.5 years (SD 4.4). The RRSO group had a more favorable CHD risk profile (higher education, more physical activity, less smoking, lower total cholesterol, higher HDL cholesterol, lower systolic blood pressure and lower BMI) and lower Framingham total score compared to controls (p<0.05). In multiple logistic regression analyses RRSO was inversely associated with Framingham 10-year risk ≥5% (Odds Ratio 0.49, 95% CI [0.34, 0.71] p<0.001). Conclusions: In contrast to expectation, women at increased risk of hereditary breast ovarian cancer had a favorable CHD risk profile after RRSO compared to age-matched controls from the general population, and RRSO was associated with lower Framingham risk score. Follow-up time, self-selection of women seeking genetic counseling, changes in lifestyle after surgery and survival bias are possible explanations of this finding.

No significant financial relationships to disclose.
Value of preoperative staging of breast cancer patients using computed tomography to detect asymptomatic lung and liver metastasis
Authors: Han W. , Kim H. , Lee J. , Lee K. , Moon H. , Ko E. , Kim E. , Yu J. , Noh D.
Published: August 2020

1105

Background: Preoperative clinical staging in breast cancer patients is important to determine the most appropriate treatment plans and to predict prognosis for individual patients. Identifying unexpected distant metastases in newly diagnosed breast cancer patients frequently alters initial treatment plans. Routine imaging studies to detect lung or liver metastasis is not indicated in patients with early and operable breast cancer. A recent study showed that routine use of chest radiograph and liver ultrasound does not provide much diagnostic benefit in early breast cancer patients. Methods: We aimed to investigate the value of preoperative computed tomography to detect asymptomatic liver and lung metastasis in breast cancer patients. We performed preoperative CT for 667 breast cancer patients to detect lung and liver metastasis among 1,636 primary breast cancer patients who had been diagnosed and treated between January 2006 and December 2007 at Seoul National University Hospital. Results: CT showed abnormal findings (suspicious of metastasis or indeterminate nodules) in 78 patients (10.5%). Among these, abnormal finding in 13 patients (1.7%) turned out to be true metastatic lesions. There was no CT-detected lung or liver metastasis in patients with T1 tumor and 4 metastases in patients with T2 tumor. There was no CT-detected lung or liver metastasis in patients with negative axillary lymph node metastasis. When patients were classified according to the AJCC staging, CT-detected true metastatic lesions were only present in stage III patients (13 out of 173 patients, 7.5%). The true metastatic lesions in lung or liver were all small sized nodules, ranging from 0.3cm to 1.2cm in largest diameters. In seven patients, the CT-detected metastatic lesions were less than 1cm which is in contrast with the previous studies. Conclusions: Our results demonstrated the lack of usefulness in performing routine CT exams to detect asymptomatic liver and lung metastasis in early breast cancer patients. Staging CT might be useful in stage III patients, since 13 (7.5%) patients were upstaged to stage IV by the use of CT.

No significant financial relationships to disclose.