bluuebunny/crossref_metadata_2025 huggingface dataset:
Leukemic blasts fail to differentiate into mature antigen presenting cells, and may hinder dendritic cell maturation in vitro and in vivo
Authors: Rosenblatt J. , Stone R. , Lenahan C. , Wu Z. , Vasir B. , Bissonnette A. , Galinsky I. , Miller K. , Levine J. D. , Joyce R. , Avigan D.
Published: March 2020

10556

Background: Dendritic cells (DC) play a key role in the development of tumor specific immune responses. Dendritic cells differentiated from leukemic blasts (LDC) are being explored as a tumor vaccine in AML. We examined the phenotypic and functional characteristics of LDC, the phenotypic characteristics of native DC in AML patients, and the effect of leukemic blasts on the phenotype of DC generated from normal donors. Methods: Leukemia blasts were isolated from peripheral blood of 24 patients with AML. LDC were generated by culturing blasts in the presence of GM-CSF, IL-4 and TNFa for 7 days. The phenotype of circulating DC1 (CD11C+/lin-) and DC2 (CD123+/ lin-) in AML patients was assessed by multichannel FACS analysis. To assess the effect of blasts on DC maturation, adherent mononuclear cells were isolated from normal donors, combined with leukemia cells in a 10:1 ratio, and cultured with GM-CSF, IL-4, and TNFa. Results: LDC demonstrate only modest expression of the costimulatory molecules CD80 and CD86 (mean expression 10% and 32%) and poorly express the maturation marker CD83 (mean expression 4%). Interferon gamma production by autologous T cells was not higher after stimulation with LDC than with blasts. LDC stimlation resulted in a 2 fold increase in both CD4+/CD25+/CD69+ (activated) and CD4+/CD25+/FOXP3+ (regulatory) T cells. Given the inability of leukemia progenitors to differentiate into phenotypically mature DC, we assessed whether leukemia cells directly inhibit differentiation of DC from normal progenitors. Expression of costimulatory molecules was decreased in DC differentiated in the presence of blasts. Mean expression of CD80, CD83, and CD86 was 16%, 2%, 83% and 49%, 10%, 99% for DCs generated in the presence or absence blasts respectively. Phenotypic characteristics of native DC in patients with AML were examined. In 3 experiments, a predominance of DC2 was seen (ratio DC2/DC1 5), and both DC1 and DC2 poorly expressed CD83 (mean expression 9% DC1, 0.9% DC2). Conclusions: LDC have phenotypic and functional deficiencies, limiting their efficacy as a tumor vaccine. Contact with leukemic blasts may inhibit DC maturation in vitro and in vivo, which may contribute to the lack of effective antitumor immunity in AML patients.

No significant financial relationships to disclose.
Role of GADD45α as a potential therapeutic target for prostate cancer
Authors: Singal R. , Ramachandran K. , Gopisetty G. , Navarro L. , Gordian E. , Manoharan M.
Published: March 2020

10566

Background: Defects in the apoptotic pathway contribute to uncontrolled cell proliferation of cancer cells and confer resistance to chemotherapeutic drugs. Understanding the mechanisms of deregulation of apoptosis related genes would enable targeted treatment methods to improve the efficacy of chemotherapy. Growth Arrest and DNA Damage inducible, alpha (GADD45a) mediates cytotoxicity of docetaxel chemotherapy. We examined the mechanism of regulation of GADD45a in prostate cancer cells and the effect of its upregulation on sensitivity to docetaxel chemotherapy. Methods: Levels of GADD45a in Du145, LNCaP and PC3 were analyzed by real time reverse transcriptase PCR and western blotting. DNA methylation was studied by bisulfite sequencing. Chromatin immunoprecipitation was used to study interaction of methyl binding proteins to GADD45 5’ sequence. Cytotoxicity after drug treatment was measured by MTT cell proliferation assay. Apoptosis assays were done by Annexin V/propidium iodide staining followed by flow cytometry. Results: Levels of expression of GADD45a in Du145 and LNCaP cells were lower than that in PC3. A 4 CpG region upstream of the proximal promoter region was methylated in Du145 and LNCaP cells. Methylation was reversed by treatment of Du145 and LNCaP cells with DNA methyl transferase (DNMT) inhibitors such as 5- Azacytidine or 5- Aza deoxycytidine leading to reactivation of GADD45a expression in these cells. This region was also frequently methylated in prostate cancer tissues. Methyl binding protein, MeCP2 was associated with the methylated 4 CpGs in Du145 and knock down of MeCP2 by transfection of MeCP2 siRNA vector in Du145 cells (Du145-MeCP2-ve) led to increased expression of GADD45a, without affecting the methylation status of the gene. Enhanced sensitivity to docetaxel was observed by upregulation of GADD45a in Du145 cells by (a) recombinant expression of GADD45a (b) downregulation of MeCP2 and (c) pretreatment with 5-Azacytidine. Conclusions: GADD45a is frequently deregulated in prostate cancer by methylation of 5’ 4 CpG region and is a potential therapeutic target for treatment of prostate cancer.

[Table: see text]
Chromosomal alterations in oligodendroglial tumors over multiple surgeries: Is progression associated with change in 1p/19q status?
Authors: Campbell B. , Horsman D. , Maguire J. , Young S. , Curman D. , Ma R. , Thiessen B.
Published: March 2020

10577

Background: There is morphologic and genotypic heterogeneity of oligodendroglial neoplasms. Tumors with loss of heterozygosity (LOH) of 1p and/or 19q are associated with increased chemo-sensitivity and survival. Despite treatment, rates of recurrence and malignant transformation are high. The pathogenesis of treatment-resistance is unknown. We aim to determine if tumour progression is associated with a proliferation of clonagens with retention of heterozygosity (ROH) of 1p, or if progressing tumours remain 1p/19q LOH. Methods: Between 1/1/2001 and 7/31/2006, 24 patients with oligodendroglial neoplasms, and possessing serial biopsies taken at diagnosis and at progression, were identified. Using PCR amplification of multiple microsatellite markers, a total of 53 tumour specimens were available for LOH analysis of 1p and 19q; 40 were also tested for 9p and 10q. Results: At diagnosis, the median age was 34 (24–66) years and 14 were male. Using the WHO criteria, 19 tumors were Grade II oligodendrogliomas or oligoastrocytomas, and 5 were high grade. The most common genomic status was 19q LOH (88%); 54% had 1p LOH. Of the 13 primary biopsies with 1p LOH, 11 had 19q LOH, 1 had 19q ROH, and 1 was 19q non-informative. A further 2 primaries were mixed 1p LOH/ROH, and 9 were 1p ROH. At progression, 10 of 11 patients with 1p/19q LOH had persistent co-deletion. The patient with 1p LOH and 19q ROH at diagnosis, had 1p/19q LOH at progression. Of the mixed 1p primaries, 1 was 1p ROH, and the other remained mixed LOH/ROH at progression. 8 of 9 primaries with 1p ROH remained 1p ROH. There was also little heterogeneity of 9p and 10q between primary and progressive tumours. Using Kaplain-Meier analysis, mean overall survival (OS) for the group was 102 (95% CI: 77–127) months. Mean progression-free survival was 52 (95% CI: 33–72) months. OS was not statistically significant between patients with 1p LOH and 1p ROH primary tumours. Conclusions: 91% of repeat biopsies of oligodendroglial tumors, demonstrated persistent 1p/19q LOH. Therefore, progression of 1p/19q LOH primary tumours is not due to a proliferating sub-group of chemo-resistant, 1p ROH clonagens. We propose that additional mutations contribute to this aggressive phenotype.

No significant financial relationships to disclose.
Prognostic role of the extent of peritumoral vascular invasion in operable breast cancer
Authors: Colleoni M. , Rotmensz N. , Andrighetto S. , Maisonneuve P. , Sonzogni A. , Pruneri G. , Casadio C. , Goldhirsch A. , Viale G.
Published: March 2020

10586

Background: Peritumoral vascular invasion (PVI) has been recently recognized as a significant prognostic indicator for women with operable breast cancer, yet the clinical relevance of the degree of PVI in patients with no or limited involvement of the axillary nodes is unknown. Methods: 2606 consecutive patients with pT1–3, pN0 (1586)-1a (1020), and M0, operated and counseled for medical therapy from 1/2000 to 12/2002 were prospectively classified according to the degree of PVI: absent (2017, 77.4%), focal (368, 14.1%), moderate (51, 2.0%) and extensive (170, 6.5%). The median follow-up was 3.8 years for disease-free survival (DFS) and 4.3 years for overall survival (OS). Results: Patients with extensive PVI were more likely be younger, to have larger tumors, high tumor grade, axillary positive nodes, high Ki-67 expression, and HER2/neu over-expression if compared with patients having less amount if PVI (p for trend, <.0001). Patients with diffuse PVI were prescribed significantly more frequently anthracycline containing chemotherapy and less endocrine therapy alone (p for trend, <.0001). In patients with node negative disease a statistically significant difference in DFS, risk of distant metastases and OS was observed at the multivariate analysis for diffuse PVI versus no PVI (Hazard Ratios: 2.11, 95% CI, 1.02 to 4.34, P<.0001 for DFS; 4.51, 95% CI, 1.96 to 10.4, P<.0001 for distant metastases; 3.55, 95% CI, 1.24 to 10.1, P=.02 for OS). Conclusions: Extensive peritumoral vascular invasion has a prognostic role in patients with axillary lymph node negative breast cancer. The extent of vascular invasion should be considered in the therapeutic algorithm in order to proper select targeted adjuvant treatment.

No significant financial relationships to disclose.
Gamma-Secretase, ErbB4 nuclear localization and neuregulin expression correlates with prostate cancer patient clinical outcome
Authors: Koumakpayi I. , Le Page C. , Karakiewicz P. I. , Diallo J. , Lessard L. , Mes-Masson A. , Saad F.
Published: March 2020

10587

Background: Membrane protein ErbB4 is a member of ErbB growth factor receptor family, which can be activated by neuregulins (NRG). Upon neuregulin activation, ErbB4 is cleaved within its transmembrane domain by presenilin γ-secratase (PSN) to release an intracellular domain that translocates into the nucleus. Although, ErbB4 ligand-dependant translocation of ErbB4 to the nucleus and its nuclear activity has been reported in breast cancer cell lines, there are few reports concerning ErbB4 nuclear localization and its clinical relevance. Here, we report for the first time the clinical relevance of ErbB4 nuclear localization, NRG, and PSN expression in prostate cancer tissues. Methods: Immunostaining using anti-ErbB4, anti-PSN2 and anti-neuregulin antibodies was done on a set of tissue microarrays (TMA) from 140 patients. The TMAs contained, 92 cores of normal prostate tissue obtained from 46 autopsy specimens from young males, 373 tumor and normal adjacent cores from 63 hormone sensitive PCa (HSPCa) patients, and 146 cores from 31 hormone refractory PCa (HRPCa) patients. Results: We found a statistically significant increase (p<0.01) in the percentage of ErbB4 nuclear localization (68.7% vs 53.2%), NRG expression (2.06 vs 1.41) and PSN2 expression (2.14 vs 1.53) when comparing cancerous tissues to normal tissue adjacent to cancer. Interestingly, a similar statistically significant increase in nuclear ErbB4 and NRG expression was observed when comparing HRPCa to HSPCa (p<0.001). In cancerous tissues, a strong correlation was found between nuclear ErbB4 and NRG expression (r=0.672), between nuclear ErbB4 and PSN2 expression (r=0.51), and between PSN2 and NRG expression (r=0.71). Nuclear ErbB4 and PSN2 inversely correlated with tumor stage and lymph node invasion. Kaplan Meier analysis of nuclear ErbB4 (p=0.030) and PSN2 expression (p=0.018) showed an inverse association with biochemical recurrence (BCR) of PCa. In multivariate analyses including these three markers and clinical parameters, only nuclear ErbB4 retained an independent prognosis value. Conclusion: Our results suggest that high nuclear ErbB4 along with increased PSN2 expression have a protective effect against prostate cancer progression and BCR.

No significant financial relationships to disclose.
Circulating tumor cells (CTCs) in peripheral blood of primary breast cancer patients
Authors: Rack B. K. , Schindlbeck C. , Hofmann S. , Schneeweiss A. , Rezai M. , Beckmann M. , Pantel K. , Schneider A. , Sommer H. , Janni W.
Published: March 2020

10595

Background: Detection of CTCs has been shown to predict decreased PFS and OAS in metastatic breast cancer, whereas only limited data has been published in the adjuvant setting. We evaluate the role of CTCs in peripheral blood at primary diagnosis and during adjuvant chemotherapy, endocrine and bisphophonate treatment within the SUCCESS-Trial (n=3,658 pts). Methods: We analyzed 23ml of peripheral blood from 1767 N+ and high risk N- primary breast cancer pts before systemic treatment. 852 of these pts have undergone follow-up blood sampling after completion of chemotherapy. The presence of CTCs was assessed with the CellSearchSystem (Veridex, Warren, USA). Briefly, after immunomagnetic enrichment with an anti-Epcam-antibody, cells were labelled with anti-cytokeratin (8,18,19) and anti-CD45 antibodies to distinguish epithelial cells and leukocytes. Results: 10% of pts with a blood sampling before systemic treatment (n=170) showed >1CTC before the start of systemic treatment (mean 13, range 2–827). While we found 2 CTCs in 5% of pts, 3% had 3–5 CTCs and 1% 6–10 and >10 CTCs each. The presence of CTCs did not correlate with tumor size (p=.07), grading (p=.30), hormonal status (p=.54) or Her2-Status of the primary tumor (p=.26). However, we observed a significant correlation with the presence of lymph node metastases (p=.015). None of 24 healthy individuals showed more than 1 CTC. Among those 852 pts with follow-up blood sampling after the completion of cytostatic treatment, 11% were CTC positive before starting systemic treatment (mean 7, range 2–166), while 7% of patients presented with >1CTC after completion of chemotherapy (mean 6, range 2–84). Of those, initially CTC positive, 10% remained positive (n=9) and 90% had a negative CTC test after chemotherapy (n=82). Of those initially CTC negative, 93% remained negative (n=711), whereas 7% returned with a positive CTC test (n=50) (p=.24). Conclusions: Our data show good feasibility of this highly standardized and easily applicable approach for the detection of CTCs in a large number of primary breast cancer patients. In a considerable number of patients, persistent CTCs can be detected after completion of cytostatic treatment. Whether this finding is prognostically relevant will have to been shown with longer follow-up of the SUCCESS-trial.

No significant financial relationships to disclose.
Preoperative sodium concentration as a possible predictor of survival in patients with renal cell carcinoma
Authors: Vasudev N. S. , Brown J. E. , Brown S. R. , Cocks K. , O'Donnell D. , Patel P. M. , Harnden P. , Banks R. E. , Selby P. J.
Published: March 2020

10601

Background: Conventional renal cell carcinoma (RCC) has a variable natural history, and determining individual prognosis is important to guide management. In this cross-sectional study of RCC, a large number of parameters including hematological and biochemical as well as traditional tumor-related factors have been assessed for their prognostic significance. Methods: Consecutive, unselected patients undergoing nephrectomy for newly diagnosed RCC were invited to participate. Primary end-points were overall survival (OS), disease-free survival (DFS) and cancer-specific survival (CSS). A subset analysis of patients presenting with localized N0 disease was planned. Univariate and multivariate Cox's proportional hazards models were used. Results: 212 patients with RCC formed the study population. Tumor-related factors including grade, stage, presence of necrosis and vascular invasion were prognostic for survival on multivariate testing, in agreement with previous reports. In multivariate analyses pre-operative serum sodium (sNa) was found to be independently and significantly associated with OS and DFS when considered as a continuous variable (see table ). In a further exploratory analysis for all patients when sNa was dichotomized into above/below the median value (139 mmol/l; normal range: 135–145 mmol/l), 3-year OS estimates above and below the median were 74%, 95% CI [64%, 85%] and 54%, 95% CI [44%, 64%]; corresponding figures for 5 years were 68%, 95% CI [54%, 81%] and 44%, 95% CI [33%, 56%]. Conclusions: We have confirmed the prognostic value of traditional tumor-related factors but, to our knowledge, these are the first data to demonstrate that pre-operative sodium concentration may be an important factor associated with survival in patients with RCC. This result suggests that sNa should be considered with established prognostic parameters in modeling survival in RCC.

[Table: see text]

No significant financial relationships to disclose.
Assessment of tumour size and its relationship to nodal involvement in multifocal and multicentric breast cancer
Authors: Rezo A. , Rodins K. , Davis A. J. , Shadbolt B. , Zhang Y. , Huynh F. , Dahlstrom J. ,   ACT & SE NSW Breast Cancer Treatment Group (ACT& SE NSW BCTG)
Published: March 2020

10602

Background: The AJCC/UICC staging system categorises tumour size in multifocal and multicentric breast cancer (MMBC) using the largest dimension of the largest focus of tumour. This under-estimates the total tumour burden and therefore may under-estimate the potential of these tumours to metastasise. This study tests the hypothesis that an aggregate measurement of tumour size in MMBC is more closely related to nodal status than the size of the largest focus. Methods: This prospective cohort study involved women with ipsilateral invasive breast cancer with known nodal status using data from the ACT & SENSW BCTG database from July 1997 to June 2004. Pathology reports were reviewed to obtain tumour size in the unifocal group and nodal status in all women. The histopathology of MMBC cases were reviewed measuring all tumour foci that were recorded as: (1) the diameter of the largest deposit (LD), (2) the aggregate diameter (AD) of all deposits, and (3) a calculation of the aggregate volume (AV) using the largest dimension of each focus. The MMBC dimensions were compared with unifocal cancers and against nodal status using a multivariate logistic regression analysis and compared against each other using a stepwise method to determine which method is most predictive of nodal involvement. Results: 795 women met the criteria for study entry: 139 (17.5%) had MMBC and 656 (82.5%) had unifocal disease. When adjusting for tumour size method of measurement in MMBC, there was strong agreement between the AD MMBC and unifocal groups in terms of nodal status (p=0.7). Conversely, there were statistically significant differences in nodal status using LD (p=0.008) and AV (p=0.02) measurements in MMBC when compared to the unifocal group. Within the MMBC group the aggregated diameter measure was most significantly related to nodal status (p=0.001). Conclusions: The results of this study confirm that the current tumour staging system in MMBC under-estimates the potential for nodal spread and that aggregate diameter is more clinically useful. Nodal status is used in this study as a surrogate for tumour behaviour. Further studies are underway to assess relationship of aggregate size of MMBC to relapse and survival.

No significant financial relationships to disclose.
Investigating tamoxifen resistance in the luminal B estrogen receptor positive breast cancer subtype: Tailoring treatment in hormone responsive breast cancer
Authors: Loi S. , Sotiriou C. , Haibe-Kains B. , Lallemand F. , Conus N. , Piccart M. J. , Speed T. , McArthur G. A.
Published: March 2020

10603

Background: We recently reported that the two estrogen receptor (ER) positive breast cancer (BC) molecular subtypes can be defined by their expression of proliferation genes using a gene expression index (GGI): the luminal A and B subtypes have low and high levels respectively (J Clin Onc, in press). When treated with adjuvant tamoxifen, luminal A tumors have a good prognosis, however the clinical outcome of the luminal B subtype was poor. This study aimed to explain the biological basis for these observations using global gene expression profiling and an in vitro model of ER+ BC. Methods: 246 ER+ BC samples from women treated with adjuvant tamoxifen monotherapy were analyzed with affymetrix gene expression arrays and evaluated using gene set enrichment analysis (GSEA). ER+ MCF-7 BC cells (control) treated with tamoxifen (TAM) and heregulin (HRG) were used to investigate molecular pathways identified using GSEA. Results: We found that a gene set suggesting ERBB2 pathway activation was significantly enriched in the luminal B subtype (p=0.02). Only 10% of samples overexpressed HER2 by immunohistochemistry, suggesting that activation of HER2 signaling pathways is independent of HER2 overexpression and may contribute to TAM resistance in this subtype. To validate this hypothesis, MCF-7 cell-lines were treated with HRG (HRG-MCF7) to create a model of ERBB2 pathway activation. HRG-MCF7 cells displayed phosphorylation of HER2/3 without HER2 overexpression. Treatment with HRG overcame TAM induced cell cycle arrest with higher S-phase fraction (p<0.01) and increased anchorage- independent colony formation (p<0.01). Gene expression profiling confirmed significant enrichment of the ERBB2 gene set (p<0.01) and higher GGI levels (p=0.02) in HRG-MCF7 cells compared with control. Conclusions: HRG-MCF7 cells may be useful as an in vitro model of the TAM resistant luminal B subtype. In this group, targeting activated HER2 signaling may be a helpful treatment strategy despite the lack of HER2 overexpression. Our data suggests that agents like lapatinib may be effective only in the luminal B and not the luminal A tumors, demonstrating the importance of stratifying by subtype in future clinical trials of ER+ disease.

No significant financial relationships to disclose.
Evaluation of gemcitabine plus carboplatin in pretreated, metastatic breast cancer patients: Final analysis of a phase II trial
Authors: Laessig D. , Vehling-Kaiser U. , Stemmler H. J. , Fasching P. , Melchert F. , Koelbl H. , Stauch M. , Scharl A. , Morack G. , Heinemann V.
Published: March 2020

1074

Background: There is an increasing proportion of metastatic breast cancer patients (pts) who have been exposed to anthracyclines and taxanes either during perioperative treatment or during intial therapy of metastatic disease. The efficacy of gemcitabine plus cisplatin has been consistently demonstrated in several trials. This study evaluates the combination of gemcitabine plus carboplatin as a treatment option for intensively pretreated breast cancer pts. Methods: Metastatic breast cancer pts were treated with gemcitabine (1,000 mg/m2 iv on days 1 and 8) and carboplatin (AUC 4 iv on day 1) in a 3-week regimen. The trial was performed as a 2-stage phase II study according to the optimal design described by Simon (p0 = 0.1, p1 = 0.3, a = 0.05, β = 0.1) with overall remission rate (according to RECIST) as the primary objective. Results: 39 pts were recruited, median age was 60 years (29–77 yrs): median Karnofsky performance status was 90% (range, 70–100%), 77% of pts were ER- and/or PR-positive, and 28% of pts presented with Her-2 overexpression (IHC 3+ or FISH-positive). 87% of pts had visceral metastasis and 79.5% had =2 sites of metastasis. All patients had received prior chemotherapy and 92% of pts had received prior anthracyclines (87%) and/or taxanes (67%). Prior endocrine therapy had been applied to 77% of patients. Median follow-up time was 13.2 months (1–27 months) during which 207 treatment cycles were documented with a median number of 5 cycles per pt (range 1–12). A CR was observed in 1 pt, a PR in 11 pts for an overall remission rate of 31% (95%-CI: 17–48%). Stable disease was documented in 12 pts (31%) resulting in a disease control rate of 61.5%. Median time to remission was 2.6 months (1.3–5.1 months), median time to progression was 4.9 months (95%CI, 2.6–6.7 months), and overall survival was 13,2 months (95% CI, 8.7–16.7 months). Grade 3–4 leucopenia (NCI-CTC) was observed in 64%, neutropenia in 51%, thrombocytopenia in 49%, and anemia in 10% of pts. Conclusion: Gemcitabine plus carboplatin is an effective and well tolerated treatment option in intensively pretreated breast cancer pts.

No significant financial relationships to disclose.
Low tumor cell detection rate in bone marrow of patients with primary breast cancer after elimination of stroma cells: Comparison of immunocytology and molecular analysis of CK19 and mammaglobin 1
Authors: Kaul S. , Aggarwal M. , Eichler A. , Sohn C. , Fersis N.
Published: March 2020

1075

Background: Rapidly self-renewing stroma cells of the bone marrow (RS cells) are MUC-1 and cytokeratin positive at a rate of 56% (475/850). Elimination of RS cells by size exclusion results in a positivity rate of 2.6% (36/1407) in primary breast cancer patients. We have used preanalytical immunomagnetic tumor cell enrichment followed by molecular detection of CK19 and Mammaglobin1 (MA) transcripts for an independent verification of the bright-field immunocytology results. Material and Methods: Mononuclear cells (2x106) from bone marrow of 1,407 patients with primary breast cancer were analysed by standardized cytospin immunocytology techniques. Disseminated tumor cells (DTC) were stained with CK8/18/19 antibodies 5D3 and A45-B/B3 using either the APAAP technique (Dako Techmate 500) or the enhanced SA-FastRed method (Ventana). Parallel DTC consisted of immunomagnetic tumor cell selection of 1x107 BM cells using the cell membrane glycoproteins EpCAM and MUC-1 as targets. CK 19 and MA transcripts were determined by conventional and quantitative real time RT-PCR. Results: Using acetic acid and periodate blocking steps before immunocytology 36 from 1407 (2.6%) breast cancer patients operated between 1999 and 2005 at the University Hspital Heidelberg were scored DCT positive using computer assisted automated (Chromavision ACIS II) picture analysis. From this group 186 bone marrow samples were used for molecular tumor cell detection. By RT-PCR analysis of 1x107 immunomagnetically purified BM cells only 6 from 186 (3.2%) samples were CK19 and MG positive. Discussion: Detection rates of tumor cells in bone marrow from non-metastatic breast cancer patients has been reported to be in the range of 0% to 100%. We show here that proliferating bone marrow stroma cells of the RS type are positive for various antigens including CK polypeptides 7, 8, 18 and 19. Elimination of these cells by size exclusion results in a tumor cell detection rate of 2,6% in 1,407 BM probes. This low detection rate could be verified in 186 BM probes by CK19 and mammaglobin1 RT-PCR. We conclude that standard immunocytology in bone marrow is severely impaired by cytokeratin expression of RS-type stroma cells.

No significant financial relationships to disclose.
A phase I trial of pegylated liposomal anthracycline and lapatinib (L) combination in the treatment of metastatic breast cancer (MBC): First evaluation of an anthracycline and lapatinib combination in the treatment of MBC
Authors: Cianfrocca M. E. , Rosen S. T. , von Roenn J. H. , Rademaker A. W. , Rubin S. D. , Friedman R. A. , Rozario C. P. , Gradishar W. J.
Published: March 2020

1079

Background: Liposomal formulations such as pegylated liposomal doxorubicin (PLD) were developed to improve the therapeutic index and overall benefit of the anthracyclines (A). L is a selective and highly competitive inhibitor of ErbB1 and ErbB2 tyrosine kinases. The combination of conventional doxorubicin and an ErbB2 targeting agent (trastuzumab), while effective, led in a randomized phase III trial to an unacceptable risk of cardiac toxicity. The combination of PLD and L however may be effective with less cardiac risk Methods: This is an open-label, phase I, dose-escalation trial of PLD at 20, 30, 45 and 60 mg/m2 IV every 4 weeks (maximum of 8 doses) and L, 1,500 mg po daily until progression in patients (pts) with MBC. EGFR and/or ErbB2 positivity was not required. Prior chemotherapy, endocrine therapy and trastuzumab were allowed however prior A use was limited to 240 mg/m2 of doxorubicin or 600 mg/m2 of epirubicin and prior EGFR targeting therapies were not allowed. Concomitant CYP3A4 inducers/ inhibitors were not allowed. A left ventricular ejection fraction (LVEF) of ≥ 50% was required. The primary objective was to evaluate the safety, tolerability and feasibility of the combination of PLD and L, particularly with respect to cardiac safety. MUGAs were performed at entry and every 8 weeks thereafter. Results: 7 patients (PLD: 20 mg/m2 - 3 pts; 30 mg/m2 - 3 pts; and 45 mg/m2 - 1 pt) with a mean age of 43 yrs (range, 33–68) have been treated for a total of 14 treatment cycles. Dose-limiting toxicity (DLT) has not been reached. One pt experienced an LVEF drop to < 50% after 4 cycles however this was accompanied by a pericardial effusion felt to be secondary to progressive disease. Adverse events observed include: grade III/IV- none; grade I/II in ≥2 pts- rash, nausea, and anorexia; grade II leukopenia, fatigue, alopecia, diarrhea, headache were also seen in 1 pt each. Preliminary response data in 4 evaluable pts reveals 1 PR, 1 SD, and 2 PD. Conclusions: In the first 7 pts treated, the combination of PLD and L has been well tolerated. One pt experienced an LVEF drop to < 50%, however this was felt likely to be disease-related. DLT has not yet been reached and accrual is ongoing.

No significant financial relationships to disclose.
Triple-negative metastatic/recurrent breast cancer: Treatment with paclitaxel/carboplatin combination chemotherapy
Authors: Chia J. W. , Ang P. , See H. , Wong Z. , Soh L. , Yap Y. , Wong N.
Published: March 2020

1086

Background: Patients with triple negative (ER, PR, Her2 negative) breast cancers do not derive benefit from Herceptin or hormonal agents. For them, conventional chemotherapy remains the only option. Recent data suggests that triple negative breast cancers (TNBC) have increased sensitivity to platinum agents. We conducted a retrospective analysis to determine the response rates of such patients treated with paclitaxel and carboplatin (TC) chemotherapy. Methods: Patients with metastatic/recurrent TNBC were included in our study. Chemotherapy administered at weekly (paclitaxel 80mg/m2 and carboplatin AUC2 on D1, D8, D15) or 3 weekly intervals (paclitaxel 175mg/m2 and Carboplatin AUC 5 on D1). ER and PR status was defined using IHC. Her 2 positive was defined by a positive FISH or 3+ IHC staining. Results: 101 patients with TNBC were diagnosed at our centre from 2002 to 2005 and of these only 23 patients had metastatic/ recurrent disease. 14 patients were treated with TC and therefore included in our analysis. Median age of patients was 53 yrs (range 36 to 68yrs). 3 patients had metastatic disease at diagnosis and 11 patients were treated for recurrent disease. Median time to recurrence for this subgroup was 20 months (range 8 to 224mths). 4 and 6 patients had prior adjuvant exposure to taxanes and anthracyclines respectively. Median prior lines of chemotherapy was 1 (range 0–4). Median number of disease sites in patients was 4. Eight patients (57%) experience a partial response (PR) with TC. 2 patients had stable disease and 4 progressive disease. Although no patients experienced complete response, 3 patients had good PR. One patient with metastatic disease had complete resolution of a large, 9cm locally invasive breast tumor after 3 cycles of TC. The second had a 90% reduction in volume of a large chest wall recurrence after 2 cycles TC. Patients who had 2 or 3 prior lines of chemotherapy also continued to show response. The median time to tumor progression was 16 weeks (range 4–28 wks). Conclusions: TC gives a high response rate in patients with metastatic/ recurrent TNBC. Patients with prior exposure to taxanes and those with large volume disease showed good response. Randomised trials are underway to compare the TC regimen with non-platinum containing doublets.

No significant financial relationships to disclose.
Phase II trial of vinorelbine (V) days 1+8 every 3 weeks and trastuzumab (T) weekly as first-line chemotherapy for HER2+ metastatic breast cancer (MBC)
Authors: Andersson M. , Nielsen D. , Sørensen P. G. , Ewertz M.
Published: March 2020

1091

Background: Weekly V and T as 1. line therapy in patients (pts) with HER2+ MBC is highly effective with reported response rates (RR) from 61–86% and median time to progression (TTP) 8.5–16 months. This study examined V given twice every 3 weeks. Methods: Eligible pts had HER2+ (IHC 3+ or FISH) MBC with measurable disease by RECIST criteria, no prior chemotherapy or T for MBC, and LVEF > 50%. They received V 35 mg/m2 i.v. days 1+8 every 3 weeks (= 1 cycle) and T weekly (4 mg/kg loading dose, 2 mg/kg thereafter i.v.) until progression or unacceptable toxicity. Planned sample size was 50, but the study was replaced by a randomised trial at 46 pts, all of them eligible. Primary endpoint was RR, secondary endpoints included TTP, overall survival (OS) and toxicity. Results: From December 2001-July 2004 46 pts were recruited at 4 centers. They received 11 (1–54) cycles (median, range). V dose was reduced due to neutropenia in 7.5% of cycles and for other reasons in 14.5%. V treatment was delayed due to neutropenia in 7.8% of cycles and for other reasons in 4.7% of cycles. Median received V dose intensity was 0,78. Cardiotoxicity: asymptomatic reduction of LVEF > 20% or >10% to below 50%: 8 pts (3 to below 50%). Toxicity grade III-IV (pts): Neutropenia: 11. Infections: 5 (one septic death). Constipation 2. 43 pts have progressed and 32 have died. RR: CR: 9 (20%) PR: 13 (28%) SD 11 (24%) (SD> 6 months 9 (20%)) PD: 10 (22%) NE: 3 (7%). Median duration of response: 18.0 months. Median TTP: 11.0 months (95% confidence interval (95%CI) 7.6–14.4). Median OS: 25.4 months (95%CI 22.7–28.0). After progression 24 received docetaxel, 14 capecitabine, 6 epirubicine, and 11 endocrine therapy. Conclusions: V given twice every 3 weeks and T weekly is a highly active and well-tolerated 1. Line treatment for HER2+MBC. Recruitment into the trial was stopped at 46 pts in favor of a phase III-trial which is ongoing in the Nordic countries comparing V (days 1+8) + T with docetaxel+T every 3 weeks.

No significant financial relationships to disclose.
Administration of further chemotherapy (CT) after capecitabine as first-line chemotherapy (CT) for metastatic breast cancer (MBC)
Authors: Debled M. , Trainaud A. , Durand M. , Floquet A. , Brouste V. , Mauriac L.
Published: March 2020

1095

Background: Based on results of randomized trials of Xeloda (X) vs CMF or taxanes (T) as first-line CT for MBC, very favorable tolerability, and patients’ (pts) preference for oral therapy, front-line X is now often used in our institution, especially for slowly progressing disease. As other CT regimens, especially T, have demonstrated improved OS, we analyzed post-X CT in these pts. Methods: Analysis was restricted to pts who began X before Jan 05, all of whom have now discontinued because of progression or toxicity. Results: In 90 eligible pts, median TTF and OS from initiation of X are 9 [95% CI 7–11] and 26 [22–30] months, respectively. 75% of pts received second-line CT. Three subgroups can be analyzed: (A) 65 pts (median age 58y, range 34–84) received further CT (median 2 regimens, range 1–6) after stopping X, including T (85% of pts), anthracycline (a/c: 42%), vinorelbine (45%), or mitomycin C (20%). 24 pts received =2 regimens including a/c and T; 7 pts received neither T nor a/c (vinorelbine, n = 6; UFT, n = 1). Median OS from initiation of X was 30 months [95% CI 27–32]. 30 pts are alive after a 16-month median follow-up since X failure; they may receive another CT; (B) 3 pts are currently receiving endocrine therapy after X failure and may receive another CT later; (C) 22 pts (24%) (median age 77y, 19 =70y) died without receiving further CT. Median OS from initiation of X was 11 months (range 0.5–23). 4 of these pts (aged 57, 71, 74, and 84y) received endocrine therapy for 2–10 months. Ability to administer further CT varies with age: of pts <75y, 89% received subsequent CT (T: 75%), whereas 37% of pts =75y received second-line CT. Conclusions: TTF and OS indicate that capecitabine is very active as first-line CT for MBC. For a large majority of pts <75y, first-line X does not compromise administration of further CT including T.

No significant financial relationships to disclose.
Gemcitabine/paclitaxel compared to gemcitabine/vinorelbine in metastatic breast cancer: An interim analysis
Authors: Bensalem A. , Bouzid K.
Published: March 2020

1097

Background: New combinations and strategies have been developed over the past 10 years including new drugs such as taxanes and gemcitabine. It is not clear whether the activity of the gemcitabine-paclitaxel (GP) combination regimen would translate into better progression-free or overall survival (OS) when compared with gemcitabine-vinorelbine (GV) especially in metastatic breast cancer. This study was conducted to evaluate the overall response rate (RR) of GP Vs GV. Secondary objectives included individual responses of GP and GV, time to progression (TTP), time to treatment failure (TTF), OS, and toxicities. Methods: Patients(pts) with histological diagnosis of stage IV or recurrent breast cancer who had PS =2 and measurable disease were randomized to receive GP (Gemcitabine: 1,250mg/m2 D1 & D8- paclitaxel: 175 mg/m2 D1, D1=D28) or GV (Gemcitabine: 1,250mg/m2 D1 & D8 - vinorelbine: 25mg/m2 D1 & D8, D1=D21). Pts received anthracycline and/or capecitabine chemotherapy in adjuvant and/or metastatic setting. Results: Of 47 patients enrolled, 24 patients were randomized to GP arm and 23 to GV arm. 72% of patients were stage IV and 28% recurrent disease. To date, all patients were qualified for safety, TTF, TTP and OS analysis. Hematologic toxicities were: Neutropenia in 23% in GP Vs 17% in GV, Anemia in 12% in GP Vs 9% in GV. Non hematologic toxicities were essentially nausea and vomiting grad 2–3 in 27% in GP Vs 31% in GV. Anti-emetic agents were administrated to decrease them. The Complete Response (CR) was 27% in GP Vs 30% in GV, the Partial Response (PR) was 23% in GP Vs 17% in GV; with an Overall Response Rate (ORR) of 50% in GP Vs 47% in GV. Median TTF in weeks was 12 in GP Vs 14 in GV. Median TTP (weeks) was 14 in GP Vs 19 in GV. Median OS (weeks) was 32 in GP Vs 50 in GV. Conclusions: In our experience, schedules incriminating gemcitabine are efficient and produce clinical benefit and there activities are very interesting. The analysis of the useful of paclitaxel or vinorelbine associated to gemcitabine demonstrates that these associations are active with no significant differences in toxicities. Therefore, the questions are what regimens, for what patients to what high responses in pre-treated metastatic breast cancer.

No significant financial relationships to disclose.
A phase II study of weekly low-dose paclitaxel plus 24-hour infusion of cisplatin as first-line chemotherapy for metastatic breast cancer (MBC)
Authors: Shen Y. , Hsu C. , Kuo S. , Lu Y. , Lin C. , Huang C. , Cheng A.
Published: March 2020

1100

Background: The dose and schedule of paclitaxel have been evolving in the past 15 years. However, the lowest effective dose of weekly paclitaxel remains unclear. In this study, we examined if a very low dose of paclitaxel can be effective in MBC. Except for the dose of paclitaxel, the chemotherapy regimen and the patient population of this study were very similar to one of our previous report (Cancer 2002; 95:2044–50). Methods: Women with chemotherapy-naïve metastatic BC, adequate performance status and organ functions were eligible. All patients received paclitaxel, 50 mg/m2 iv 1 hr on D1, 8 and 15, and cisplatin, 40 mg/m2 iv 24 hrs on D1 and 8, every 4 weeks. Patients continued this low-dose regimen until progressive disease (PD) or prohibitive toxicities occurred. Patients who had PD or stable disease (SD) without clinical benefit were shifted to paclitaxel 80 mg/m2 iv 1 hr on D1, 8 and 15, and cisplatin, 40 mg/m2 iv 24 hrs on D1 and 8, every 4 weeks. The primary endpoint was overall response rate (ORR). Simon’s optimal two-stage design (P0, P1,a, and βare 20%, 40%, 0.05, and 0.2, respectively) was adopted. More than 4 objective responses were needed in the first 13 patients, and a total of 43 patients will be enrolled. Results: After a median follow-up duration of 16.5 months, 27 patients were evaluable for response and toxicity. Their median age was 52.7 years (range: 30.1∼69.4). Twenty patients were hormone receptor (+), while 6 patients were Her-2/neu (+). A total of 121 cycles of protocol treatment were administered as of Dec. 2006. There were 0 complete response and 14 partial responses, for an ORR of 51.9%. Nine of 14 responders remain progression-free. The PFS was 9.6 months (95% of CI: 5.53∼13.67%) and the OS not reached. Grade 3–4 anemia, neutropenia, liver toxicity, thrombocytopenia, and skin toxicities were reported in 8, 6, 2, 1, and 1 patients, respectively. Neurotoxicity was common (22/27), but all were grade 1 or 2. Conclusions: The preliminary results of this study suggest that weekly low-dose paclitaxel is as effective as conventional-dose paclitaxel, and the treatment-related toxicities were more acceptable.

No significant financial relationships to disclose.
The role of transvaginal ultrasound in early diagnosis of tamoxifen-related endometrial cancer: An Italian experience
Authors: Martella F. , Giannessi P. G. , Coltelli L. , Giuntini N. , Safina V. , Falcone A.
Published: March 2020

11021

Background: women treated with tamoxifen for ER-positive early breast cancer are at a two to seven fold increased risk of endometrial cancer. Even though published data fail to support the use of transvaginal ultrasound (TVU) for screening of endometrial cancer, this is still a very common clinical practice in this subset of patients (pts). Methods: we have conducted a retrospective analysis to investigate the value of TVU in early detection of tamoxifen-related endometrial cancer. The screened population consists of pts treated with adjuvant tamoxifen in our institution from January 1999 up to December 2003 receiving a TVU annually or in case of gynaecologic symptoms. Results: 491 evaluated pts performed a total of 1634 TVUs in asymptomatic conditions. FIGO stage I endometrial cancers have been diagnosed in 3 patients (0.32%) who are still alive after total hysterectomy. A vaginal bleeding anticipated the examination in 33 women (3.2%) and represented the first symptom in two cases of tumor. Only one endometrial cancer has been detected with the screening procedure. Median increase of endometrial thickness has been 7.6 mm (range 1–34 mm) and those patients with abnormal images at TVU underwent an hysteroscopy with endometrial biopsies (169) resulting in a benign histology (polyps, cystic atrophy, hyperplasia) in most cases. Conclusions: therefore we have performed more than 1500 transvaginal ultrasound to detect only one asymptomatic cancer so we agree with literature in supporting that women receiving tamoxifen should undergo only an annual gynaecologic examination reserving the TVU to patients with vaginal bleedings or discharges.

No significant financial relationships to disclose.
A phase I/II study of imatinib and docetaxel as neoadjuvant therapy in locally advanced breast cancer
Authors: Haley B. B. , Ashfaq R. , DeHaas M. , Ramaswami A. , Sikder K. , Tripathy D.
Published: March 2020

11039

Background: Imatinib mestylate(Gleevec,G) is a tyrosine kinase inhibitor of platelet derived growth factor receptor(PDGFR) and KIT(CD117). In breast cancer(BC),PDGFR signaling regulates tumor interstitial fluid pressure(IFP). PDGFR inhibition lowers IFP enhancing tumor drug delivery and chemo effect. KIT’s role in BC growth is unclear.PDGFR and KIT signal pathways regulate proliferation and cell cycle progression.Receptor blockade may enhance anti-tumor activity combined with chemo.This study evaluated tolerability,toxicity and efficacy of imatinib and docetaxel and treatment changes in biomarker expression and downstream mediators. Methods: Eligibility:Total 12 women, invasive LABC(T2–4,No-2,Mo)IHC + for KIT and/or PDGF, ECOG 0–2,bidimensional disease.Treatment:Assigned to 1of 3 G dose levels(400,600,800mg) taken daily 2 weeks then combined with D30mg/m2 IV weekly x3 Q28 days. Three completed G(400mg D) and protocol modified to G600 and 800mg dose for 1 week then combined with D30mg/m2 IV weekly x12. Tumor(T)measured pretreatment and at definitive surgery at conclusion of G+D.T biopsy done preG, postG and at end of G+D for tissue markers. Three treated G400, 5G600 and 4G800mg dose. Results: 12 patients median age 51 (range 39–62), ECOG PS O, 4 pre/8 post menopausal treated. Clinical pretreatment (C-PRx) tumor size (TS)median 6cm (range 3- 12cm).C-PRx node status 7cN0 and 5cN1.PRx x-ray TS median 4.5 cm (range 2.5–8.5cm). Response:OR 25% 1 near CR+2 PR(3/12) at 400 and 600mg dose levels.Two SD(G800mg),3 progressed/off study,2 withdrew/toxicity(G3 rash/edema and G4 N/V), 2 NR.Surgery:5PM and 7MRM. Toxicity:Non- hematologic:nausea(1G4/G400mg),vomiting(1G4/G600mg),diarrhea(1G3/G800mg),fatigue(2G3/G800mg),myalgia(1G3/G800mg),rash(1G3/G600m g) and hypophosphatemia (2G3/G600mg). Hematologic toxicity mild neutropenia/anemia G1/2.One hospitalized for G3 odonophagia/stomatitis,G800mg.Dose Delay:2 missed 1 week of D+G (G600mg),2 missed 2 weeks of D+G (1G600mg and 1G800mg). Conclusion: Imatinib and docetaxel in neoadjuvant LABC is feasible and tolerable with low G3/4 toxicity and acceptable activity.G400mg dose best tolerated. Tissue IHC and molecular profiles to be reported separately.
Oblimersen (O) in combination with docetaxel (T), adriamycin (A), and cyclophosphamide (C) as neoadjuvant systemic therapy (NST) in primary breast cancer (PBC): Final results of a multicentric phase I study
Authors: Rom J. , Schlehe B. , Marmé F. , Schuetz F. , Eichbaum M. , Sinn H. , Buechele T. , Sohn C. , von Minckwitz G. , Schneeweiss A.
Published: March 2020

11047

Background: The Bcl-2 downregulator O in combination with standard cytotoxic chemotherapy leads to synergistic antitumor effects in preclinical trials. To evaluate maximum tolerated dose (MTD), safety and preliminary efficacy of O in combination with TAC as NST in PBC we performed a multicentric phase I study. Methods: Previously untreated patients with PBC T2–4a-c N0–3 M0 received escalating doses of O as a 24 hour continuous infusion on day (d) 1–7 in combination with T 75mg/m2, A 50mg/m2 and C 500 mg/m2 on day 5 followed by five three weekly cycles of TAC without O. Results: A total of twenty-eight patients with a median age of 50 years were enrolled. 61% of tumors were T2, 43% grade 3, 68% ductal-invasive, and 64% hormone receptor positive. Patients received O in a dose of 3 mg/kg/d (cohort I, 9 patients), 5 mg/kg/d (II, 9 patients), and 7 mg/kg/d (III, 10 patients), respectively. No dose-limiting toxicity occurred. Most common adverse events [all National Cancer Institute-Common Toxicity Criteria (NCI-CTC) grade = 2] were fatigue, nausea, alopecia, headache and flue-like symptoms observed in 78% (I), 89% (II), and 90% (III) of patients, respectively. Following OTAC the most severe hematological toxicity was neutropenia (NCI-CTC grade 1–2/3/4) which developed in 0/0/56% of patients (I), 11/0/56% of patients (II), and 20/20/50% of patients (III), respectively. At least 50% reduction in tumor volume was oberved in 14 patients. Five patients (18%) achieved a pathologic complete response (3 patients in cohort I, 2 patients in cohort II). Conclusions: O up to a dose of 7 mg/kg/d given as a 24 hour infusion on day 1–7 can be safely administered in combination with standard TAC on day 5 as NST in patients with PBC. The preliminary efficacy warrants further evaluation of this combination in a randomised trial.

No significant financial relationships to disclose.