bluuebunny/crossref_metadata_2025 huggingface dataset:
Superiority of melphalan-prednisone (MP) + thalidomide (THAL) over MP and autologous stem cell transplantation in the treatment of newly diagnosed elderly patients with multiple myeloma
Authors: Facon T. , Mary J. , Harousseau J. , Huguet F. , Berthou C. , Grosbois B. , Anglaret B. , Azzedine A. , Rodon P. , Peny A.
Published: March 2020

1

Background: The standard MP regimen remains the reference treatment for elderly patients (pts) with multiple myeloma (MM). In May 2000, we initiated the IFM 99–06 trial, for pts aged 65–75 y, comparing MP (12 courses at 6 weeks intervals) to MP-THAL (MP plus THAL at the maximum tolerated dose, but ≤ 400 mg/day) and a MEL100-based treatment (VAD×2, CTX 3g/m2, and 2 courses of MEL100 mg/m2). Methods: IFM99–06 was planned to enroll 476 evaluable pts, whose treatment allocation followed a 3 (MP), 2 (MP-THAL), 2 (MEL100) randomization scheme. The primary end-point was overall survival (OS). Secondary end-points were response to treatment and progression-free survival (PFS). Two interim analyses were planned and reviewed by a Data Safety Monitoring Board (DSMB) independent of IFM. At the second interim analysis, the DSMB suggested that a third interim analysis be performed with a date of point on May 1, 2005. Results: At this time, 436 pts had been enrolled, 191, 124 and 121 in MP, MP-THAL and MEL100 groups, respectively. The median (se) follow-up time was 32.2 (1.8) months (mo.). Median (se) PFS times were 17.2 (1.5), 29.5 (3.6) and 19.0 (1.3) mo. in MP, MP-THAL and MEL100 groups, respectively. The PFS time was significantly longer in the MP-THAL group than in the MP group (RR=2.4, P<0.0001), but no significant difference was noted between MP and MEL100 groups (RR=1.2, P=0.12). There was a clear advantage in favor of MP-THAL vs MEL 100 (RR=2.0, P=0.0001). The PFS advantage in favor of MP-THAL translated to a significant benefit in terms of OS. Median (se) OS times were 30.3 (5.8) mo. (86 deaths), not reached at 56 mo.(34 deaths) and 38.6 (3.0) mo. (54 deaths) in MP, MP-THAL and MEL100 groups, respectively. The OS time was significantly longer in MP-THAL group than in MP group (RR=1.9, P=0.0008), but not significantly different between MP and MEL100 groups (RR=1.1, P=0.55). MP-THAL was also superior to MEL100 (RR=1.7, P=0.014). Conclusion: Since these results show the superiority of MP-THAL, enrollment was stopped. The final analysis will be presented at the meeting. MP-THAL should be, at the present time, the reference treatment for newly diagnosed MM pts ineligible for high-dose therapy.

No significant financial relationships to disclose.
Pharmacogenomic predictor of complete response to preoperative paclitaxel and 5-fluorouracil, doxorubicin, cyclophosphamide chemotherapy in breast cancer
Authors: Pusztai L. , Hess K. R. , Anderson K. , Valero V. , Ibrahim N. K. , Mejia J. A. , Vidaurre T. , Gomez H. L. , Hortobagyi G. N. , Symmans W. F.
Published: March 2020

10000

Background: We developed a multi-gene predictor of pathologic complete response (pCR) to preoperative weekly paclitaxel and 5-fluorouracil, doxorubicin, cyclophosphamide (T/FAC) chemotherapy and assessed its predictive accuracy on independent cases. Methods: 133 patients with stage I-III breast cancer were included. Pretreatment gene expression profiling was performed with Affymetrix U133 A chips on fine needle aspiration specimens. We developed predictors of pCR from 82 cases and assessed accuracy on 51 independent cases. Results: Overall pCR rate was 26% in both cohorts. In the training set, 56 probes (49 genes) were identified as differentially expressed between pCR versus residual disease at a false discovery rate of 1%. We examined the performance of 780 distinct classifiers (set of genes + prediction algorithm) in full cross validation. Many predictors performed equally well. A nominally best 30-probe set DLDA classifier was selected for independent validation. It showed substantially higher sensitivity (92% vs 61%), negative predictive value (96% vs 86%) and better AUC (0.877 vs 0.811) than a predictor including clinical variables (age, ER, grade). The combination of genomic and clinical information yielded the best model, AUC=0.883, sensitivity 92%, specificity 71%, PPV 52%, NPV 96%. In 31 samples, RNA was hybridized in replicate with resulting predictions that were 97% concordant. Conclusion: A 30-probe set pharmacogenomic predictor alone or in combination with ER and grade predicts response to T/FAC chemotherapy with high sensitivity and negative predictive value. This test correctly identified all but one of the patients who achieved pCR (12/13) and all but one of those who had residual cancer (27/28).

No significant financial relationships to disclose.
Identification of integrin as potential molecular markers in human glioblastoma multiforme: Microarray analysis guided by contrast-enhanced MRI
Authors: Yang Y. , Chia Y. , Rubin D. , Harsh G. , Atlas S. , Bednarski M. , Guccione S.
Published: March 2020

10007

Background: Glioblastoma multiforme (GBM) is a primary brain tumor with poor prognosis and low survival rate. It is characterized as a highly infiltrative and vascular tumor, with morphologic heterogeneity including necrosis regions. We have used contrast enhanced (CE)-MRI to identify regions that differ in vascular permeability as delineated by the small molecule contrast agent Magnevist. Non-necrotic areas with and without contrast enhancement were sampled in the OR. The expression profiles for integrin family genes were investigated and validated using IHC. Genes encoding for integrin subunits with high expression levels in these regions of the tumor were identified and are presented here. Methods: Patients diagnosed with GBM, without any prior procedures were scanned on a GE 1.5T MRI scanner using standard T1- and T2-weighted pulse sequences and Gd(DTPA) as contrast agent. Samples from regions with vascular permeability to the contrast agent (contrast-enhancing, CE) and vascular regions that are not permeable to the contrast agent (non-enhancing, NE) were collected for gene expression profiling using the Affymatrix oligonucleotide microarray system. Genomic data was normalized at probe-level, and analyzed using a modified t-test, Significance Analysis of Microarray (SAM). Significant expression levels were identified. Immunohistochemical staining was used to validate protein expression of selected genes of interest. Results: Tissue samples from the CE and NE regions of 13 patients reveal significantly distinct gene expression patterns. In specific, genes in the integrin family such as integrin αIIb, αV and β3 show high expression levels in the CE region. Immunohistochemical staining confirmed correlation of protein expression patterns with the observed genomic profile. Conclusions: Combination of CE-MRI using the clinical contrast agent and microarray analysis can reveal spatial genomic profiles correlated with imaging features associated with increased vascular permeability, vessel density, and areas of fluid accumulation. The identified genes from the integrin family can be potential molecular targets, and can be used for molecular imaging and therapeutic purposes.

No significant financial relationships to disclose.
Microsatellite instability but not thymidylate synthase is a prognostic variable in primary colon cancers from patients treated in 5-FU-based adjuvant studies
Authors: Sinicrope F. A. , Rego R. L. , Halling K. C. , Foster N. R. , Sargent D. J. , Laplant B. , French A. J. , Allegra C. J. , Laurie J. A. , Goldberg R. M. , Thibodeau S. N.
Published: March 2020

10019

Background: Colon cancers with microsatellite instability (MSI) display resistance to 5-fluorouracil (5-FU), and in vitro resistance can be reversed by restoring DNA mismatch repair proficiency. 5-FU inhibits the thymidylate synthase (TS) enzyme and TS may predict clinical outcome after 5-FU-based chemotherapy. To define molecular predictors of prognosis, we analyzed TS, p53, chromosome 17p allelic imbalance (AI), and patient survival stratified by MSI status. Methods: Primary colon carcinomas from patients enrolled in five 5-FU-based adjuvant therapy trials were analyzed for MSI and 17p AI using 11 microsatellite markers (MSI-H: ≥ 30% of the loci demonstrating instability). For 17p AI, markers included D17S261 and TP53 at or near the p53 locus. Expression of DNA mismatch repair (hMLH1, PharMingen; hMSH2; Oncogene), TS (TS106, Zymed), and p53 (D07, Novacastra) proteins were analyzed by immunohistochemistry. Correlations between markers and associations with overall survival (OS) were determined. Patients were censored at 5 years for DFS and at 8 years post study randomization for overall survival (OS) data. Results: Of 320 Dukes’ stage B2 and C cancers studied, 60 of 320 (19%) were MSI-H. TS expression variables (intensity, extent, weighted score) were similar in MSI-H and MSI stable/low frequency (MSS/MSI-L) cancers; similar results were found using DNA mismatch repair (dMMR) proteins. MSI-H tumors had lower stage (p= 0.0007), fewer metastatic lymph nodes (p= 0.004), and improved OS (vs. MSS/MSI-L tumors; p= 0.01). Loss of dMMR proteins was also associated with better OS (p= 0.006). None of the TS variables were prognostic for OS. Histologic grade (p= 0.0008) and nodal status (p= 0.0002) were associated with OS in contrast to 17p LOH or p53. Only MSI status or dMMR, histologic grade, and tumor stage were independent markers for OS. Conclusions: MSI-H tumors show earlier stage at presentation and better stage-adjusted survival rates. MSI status and TS expression were unrelated and TS was not prognostic, suggesting that TS levels cannot explain therapeutic resistance to 5-FU reported in MSI-H colon cancers.

[Table: see text]
Determination of microvessel density by real-time PCR in esophageal cancer: Correlation with histological methods, angiogenic growth factor expression and lymph node metastasis
Authors: Loges S. , Claussen H. , Reichelt U. , Bubenheim M. , Erbersdobler A. , Schurr P. , Yekebas E. , Izbicki J. , Bokemeyer C. , Fiedler W.
Published: March 2020

10053

Background: Neoangiogenesis and lymphangiogenesis represent prognostic factors in human malignancies. Determination of microvessel density (MVD) by immunohistology is labor-intensive and subject to interobserver variability. We evaluated real time PCR to quantify MVD in primary tumor samples from patients with esophageal cancer. Methods: We performed real-time PCR analyzes of endothel-specific (VE-Cadherin, P1H12, VEGFR-2, tie-2), lymphendothel-specific (Prox, LYVE, VEGFR-3) antigens and of angiogenic growth factors (VEGF-A, VEGF-C, VEGF-D, Ang-1, Ang-2) in primary esophageal carcinoma tissue of 54 patients. These results were compared to MVD determined immunohistochemically by CD31 staining. Results: For validation, MNC samples spiked with HUVECs were analyzed by qPCR for VE-CAD and P1H12 yielding a linear correlation (r=0.99 and 0.96 respectively). Expression of endothelial markers was highly correlated in tumor samples, e.g. CD144 with CD146 τb=0.451, CD144 with VEGFR-2 τb =0.744 and CD144 with tie-2 τb=0.684 (p for all comparisons < 0.0001). QPCR results were compared to MVD determination by CD31 staining in a subgroup of 33 patients. The highest association between both methods was found for CD144 (τb=0.258, p=0.0379) and VEGFR-2 (τb=0.222, p=0.0745) indicating that immunohistology and qPCR yield comparable results. MVD was significantly linked to the expression of VEGF-A, -C,-D and Ang1 and Ang2 (p for all comparisons <0.0001). We analyzed expression of lymphendothelial cell antigens Prox, LYVE and VEGFR-3 for quantification of lymphatic vessels. A close association between the expression of different lymphendothelial factors was seen (LYVE with Prox τb=0.334, p=0.0021, LYVE with VEGFR-3 τb=0.450, p=0.0150). MVD and lymphvessel density was not linked. Lymph node metastases detected on surgical specimen were associated with MVD determined immunohistologically (p=0.003)or by qPCR (p=0.048) and to VEGF-C expression (p=0.04). Conclusions: QPCR analysis of CD144 and VEGFR2 represents a novel tool for quantification of MVD in tumor samples. Expression of VEGFR2 and VEGF-C is associated with lymph node metastasis in patients with esophageal carcinoma.

No significant financial relationships to disclose.
A new genomic approach to describe the natural history of node negative breast cancer patients
Authors: Saghatchian M. , Lidereau R. , Delaloge S. , Koscielny S. , Kauffmann A. , Michiels S. , Mathieu M. , Brain E. , Dessen P. , Lazar V. , Tursz T.
Published: March 2020

10058

Background: Breast cancer is a heterogeneous and complex disease beyond good/bad prognosis groups. Previous studies identified prognostic tools that are undergoing inter-group validation. Our present study aims at better defining pre-malignant vs. malignant lesions and tumors that will give local vs. distant metastases (DM) or early vs. late DM. Methods: We selected 3 groups of patients (pts) in a consecutive series of node-negative breast cancers with a very long follow-up (>10 years (y)) and available frozen tumor samples. Pts who did not receive any adjuvant treatment were selected: 60 with no relapse after 10 y, 29 with early DM before 5 y and 18 with late DM after 5y. Genomic profiling on 44K Agilent pangenomic arrays taking as reference non-relapsing pts (after 10y) or normal breast (Clonetech) allowed us to select specific genes related to DM occurrence or tumorigenesis. Random permutations were performed to assess the statistical significance of our prediction accuracy. Results: 1- we identified a 141 gene-profile and related genes distinguishing tumors with early DM from tumors with late DM (71% prediction accuracy). 2- Similarly, we identified a 285-gene signature for late DM vs. no relapse at 10 y (80% prediction accuracy). Genes were further classified according to family clusters allowing description of genes involved in DM occurrence. 3- a total of 435 genes were also significantly over-expressed in at least 90% of 148 tumours (compared to normal breast). Among those, a selection of the most significant genes were analysed for RT-PC expression throughout the various steps of tumour development (normal breast, benign tumors, in situ carcinomas, invasive carcinomas). Among the 15 most upregulated genes listed, 3 unknown genes appear, that are overexpressed in 98% of the tumors with an average fold change > 11 and are currently explored. Discussion: In this unique series of untreated node-negative breast cancers, identification of molecular profiles of early and late DM could be useful in better early prediction of breast cancer outcome. Based on genomic analysis, a molecular gene based classification of progression from benign tissue to aggressive tumors allows to dissect pathways towards malignancy and to identify early diagnostic markers or targets for prevention.

No significant financial relationships to disclose.
Neuregulin (NRG) expression modulates clinical response to trastuzumab in patients with metastatic breast cancer (MBC)
Authors: De Alava E. , Abad M. , Rodriguez C. A. , Montero J. C. , Serrano E. , Ocana A. , Torres A. , Ruiz M. , Cruz J. J. , Pandiella A.
Published: March 2020

10069

Background: ErbB2 overexpression is the major determinant of response to trastuzumab (Herceptin, H). Recently, some authors have reported the potential role of the expression of NRG on ErbB2 activation in breast cancer cells, and its consequences in response to treatment with H. In the present study we analyze the relationship between response to H and expression of ErbB2 and NRG in patients (pt) with MBC. Methods: Data and frozen tissue samples from 30 consecutive pt with MBC and positive ErbB2 at diagnosis [Immunohistochemistry (IHC)] were collected. All pt were treated with an Herceptin-based regimen. Central Pathologic review of ErbB2 expression was performed in all samples (Herceptest and FISH). NRG expression was studied by IHC on paraffin embedded material and by Western blotting on frozen tissue using an antibody raised against the intracellular domain of NRG. NRG expression by IHC was evaluated using a semiquantitative scoring system that assesses both extension and intensity of cytoplasmic staining. Correlation between ErbB2 and NRG expression and its influence in response to H was analyzed. Characteristics of pt: Median Age: 54; Median mts sites: 2; Treatment: H alone:1 pt; H+Taxol:15 pt; H+Taxol+Carbo: 4 pt; H+Taxol+Lip.Doxorub:2 pt, H+Vinorelbine:7 pt; H+CDDP:1 pt. Results: ORR: 76% (CR:23%, PR:53%). Median TTP: 7.8 m. After central Pathologic review, a total of 20 tumors showed ErbB2 amplification by FISH and 19 of them were Herceptest 3+. A positive correlation was observed between ErbB2 amplification and NRG expression. All complete responses (n=7) were seen in pt with ErbB2 amplification. Interestingly, in the group of tumors without ErbB2 amplification (n=10), 7 had high NRG expression levels, as assessed by IHC. Six out of these 7 tumors having high levels of NRG expression, exhibited partial responses. NRG expression, in turn, did not have impact on response among ErbB2 amplified tumors. Conclusions: These preliminary results suggest that responses to H regimens in pt with MBC can be seen in pt lacking ErbB2 amplification in presence of high levels of expression of transmembrane ligand NRG. This suggests that the group of pt that may benefit from treatment with H could be broader than currently established. A confirmatory study is ongoing in a larger series.

No significant financial relationships to disclose.
Immunization with MUC1/X protein enhances cDNA immunization in generating anti-MUC1 alpha/beta junction antibodies that target cancer cells
Authors: Rubinstein D. B. , Ziv R. , Karmely M. , Leitner O. , Wreschner D.
Published: March 2020

10072

Background: MUC1, a glycoprotein highly expressed in epithelial malignancies including breast, prostate, and ovarian, and on the malignant cells of multiple myeloma has generated considerable interest as a tumor marker and target for tumor killing. The most intensively studied MUC1 protein is a type I transmembrane protein (MUC1/TM) which is proteolytically cleaved soon after synthesis into α and β subunits which bind in a strong non-covalent interaction. Almost all antibodies generated to date against MUC1 recognize epitopes within the highly immunogenic tandem-repeat-array. A major shortcoming in use of such antibodies is the fact that the tandem-repeat-array-containing part of MUC1 is shed from the cell surface into the circulation. Soluble, shed MUC1 sequesters circulating anti-tandem-repeat-array antibodies, limiting their ability to reach targeted MUC1-expressing cells. Antibodies to MUC1 epitopes tethered to the cell surface would likely be more effective therapeutic agents. Despite efforts in recent years, such antibodies have remained elusive; generation of anti-cell antibodies requires characterization of cell-bound epitopes. The junction of the MUC1 α-subunit binding the membrane-tethered β-subunit provides such an epitope. Methods: By use of a novel protocol, entailing immunization with MUC1/TM cDNA and boosting with MUC1/X protein, a MUC1 isoform lacking the tandem-repeat-array, we generated monoclonal antibodies designated DMC209 which recognize the MUC1 α/β junction. Results: DMC209 is exquisitely unique for the target site; all amino acid mutations which abrogate MUC1 cleavage also abrogate DMC209 binding. Additionally, DMC209 binds the MUC1 α/β junction on cell-tethered tandem-repeat-array-containing MUC1 (MUC1/TM) on breast and ovarian cancer, and on myeloma cells. Conclusion: DMC209 is likely to have clinical application by targeting MUC1-expressing cells directly, and as an immunotoxin conjugate. Moreover, the novel high-titer immunization procedure used in generating DMC209 can be used to generate anti-MUC1 α/β junction antibodies acting as ligand and, analogously to herceptin, may have direct cytotoxic activity.

No significant financial relationships to disclose.
Predicting survival and metastasis in soft tissue sarcoma: A gene microarray study using RNA derived from archival paraffin-embedded tumours
Authors: Linton K. M. , Hey Y. , Jeziorska M. , Wilson C. L. , Miller C. , Saunders E. , Pepper S. , Freemont A. , Radford J. A.
Published: March 2020

10074

Background: The genetic determinants of survival and metastasis in soft tissue sarcoma (STS) are poorly understood and progress in this field has been limited by the rare nature of STS and the need for fresh/frozen tissue (FT) for gene microarray analyses. Our objective was to determine whether valid gene microarray data can be obtained from archival formalin-fixed paraffin-embedded tumours (FFPET) in order to retrospectively identify prognostic STS gene signatures and potential molecular targets from patients with known clinical outcome data. Methods: Total RNA was extracted from macrodissected viable tumour tissue for 34 FFPET (14 liposarcomas, 11 leiomyosarcomas and 9 synovial sarcomas) and two paired FFPET and FT primary leiomyosarcomas (modified Optimum kit, Ambion, for FFPET RNA extraction and Trizol, Invitrogen, for FT RNA extraction). One-cycle labelled cRNA was hybridised to Affymetrix U133 Plus 2 microarrays, and strict QC protocols used to identify 19 arrays for further analysis using R and BioConductor. Log rank regression and Kaplan-Meier plots of disease-specific survival were performed to identify genes predictive of survival and/or metastasis. Results: Similar fold changes in gene expression were obtained for paired cRNA samples, although the number of genes called present was lower for cRNA from FFPET. Five hundred genes (selected from FFPET cRNA arrays) were discriminatory for survival (p=0.0006 to 0.0088) across all STS subtypes and correctly assigned 17/19 cases by leave-one-out cross validation. Many are known tumour prognostic genes, significant for survival, metastasis, invasion, angiogenesis and apoptosis. Twenty-five novel and known candidate genes (with 3–10 fold differential expression) were selected for validation on 19 test and 60 independent cases by RT-PCR for relative gene expression and immunohistochemistry for protein detection. Conclusions: We have shown that prognostic information can be derived from archival FFPET, permitting the identification of candidate prognostic genes and therapeutic targets in STS, and opening the way for studies in other tumours where FT is unavailable.

No significant financial relationships to disclose.
Estrogen receptor β represses breast tumor growth and angiogenesis in vivo
Authors: Hartman J. , Lindberg K. , Inzunza J. , Wan J. , Ström A. , Gustafsson J.
Published: March 2020

10101

Background: Estrogens are well known stimulators of breast cancer cell growth in vitro as well as in vivo. Two different estrogen receptors exist, namely estrogen receptor (ER) α and β. ERα mediates the proliferative effect of estrogen in breast cancer cells and we have earlier shown that ERβ inhibits cell-cycle progression in vitro. Estrogens are well known stimulators of in vivo breast cancer cell growth as well as angiogenesis, and the effect is mediated through ERα. The function of ERβ in this context is not well understood. Methods: We have used ERα-positive T47D breast cancer cells stably transfected with a Tet/Off regulated ERβ expression vector system. The ERβ-inducible tumor cells are studied in vitro as well as in vivo. Results: By transplanting ERβ-inducible breast cancer cells into SCID-mice, we show that ERβ inhibits tumor growth and reduces the volume of established tumors. Furthermore, we show by immunohistochemistry, that the number of blood microvessels in the tumor periphery is decreased by ERβ expression, counteracting the well-known pro-angiogenic effect of ERα. By Western blot analysis on tumor extracts, we show that the concentration of the important pro-angiogenic growth factors VEGF and bFGF, normally expressed by breast tumor cells, is decreased in the ERβ-expressing tumors compared to the normal tumors. To exclude that the observed anti-angiogenic effect is just a result of reduced tumor growth, we incubated Tet/Off regulated ERβ expressing cells in vitro, during non-hypoxic conditions. We found that the expression of ERβ leads to decreased expression of VEGF and PDGFβ at the mRNA and protein-levels. In transient transfection assays, we found estrogen-ERα mediated up regulation of VEGF, PDGFβ and bFGF-promoter activities in T47D cells, and these activities were all suppressed following co-transfection with an ERβ-expression vector. Conclusions: We conclude that ERβ inhibits growth factor expression at transcriptional level in breast cancer cells; taken together, our data indicates that ERβ inhibits growth and angiogenesis of tumors formed by T47D breast cancer cells. This makes ERβ an interesting therapeutic target in breast cancer and perhaps treatment with the newly designed ERβ-selective ligands might work as a new anti-proliferative and anti-angiogenic therapy.

No significant financial relationships to disclose.
Heterogeneity of epidermal growth factor receptor (EGFR) expression and variation in immunohistochemistry (IHC) testing may affect access to EGFR-targeted therapy in patients with advanced colorectal cancer (CRC)
Authors: Beer M. W. , Ung C. , Bacus S. S. , McCollum A. D. , Ervin-Haynes A. , Schinagl R. , Youssoufian H. , Rowinsky E.
Published: March 2020

10104

Introduction: Cetuximab, an IgG1 MAb directed at the EGFR, has demonstrated benefit in pts with CRC and is currently indicated in EGFR expressing CRC. However, previous reports show heterogeneous EGFR expression between anatomic sites and suggest that results from IHC testing vary between labs. To assess this variability and the resultant impact on pt access to EGFR-targeted therapy, we compared EGFR results reported to clinicians by hospital and reference labs with those obtained at a single central lab. Methods: Formalin-fixed, paraffin-embeded CRC specimens submitted through the Targeted Diagnostics Advocacy Program (tdap) were assayed using the Dako EGFR pharmDx kit and scored using the FDA-approved package insert. If the initial specimen tested EGFR negative (−), additional specimens were requested for assay. Prior EGFR results, lab, and specimen ID were documented through review of anatomic pathology reports. Results: Of 332 evaluable CRC pts tested, 245 pts (74%) were previously reported as EGFR(−). After retesting, 167 (68%) pts were found to be EGFR The originally tested tumor block was retested in 82 pts and 41 pts (50%) were found to be EGFR (+). Retest results were similar for both hospital and reference labs. Of 66 pts with >1 specimen from the same anatomic site tested, 21 (32%) were discordant (at least 1 (+) and one (−) result). Results from different anatomic sites were compared for 65 pts, 34 (52%) of which were discordant (+). Overall, 231 pts (70%) tested EGFR(+). Conclusion: This study shows marked heterogeneity in EGFR expression by IHC both within and across anatomic sites from the same pt as well as variability in both hospital and reference pathology labs. This variability undoubtedly has an adverse impact on pt access to EGFR-targeted therapy. The impact of EGFR heterogeneity may be reduced through the assay of multiple specimens.

[Table: see text]

[Table: see text]
Correlation of mammographic breast density with Ki-67 expression in benign breast epithelial cells obtained by random periareolar fine needle aspiration of high risk women
Authors: Khan Q. J. , Kimler B. F. , Smith E. J. , O’Dea A. P. , Sharma P. , Fabian C. J.
Published: March 2020

1011

  >Background: Known risk factors for breast cancer development include elements incorporated into the Gail risk model, mammographic breast density and cytologic atypia detected by Random Periareolar Fine Needle Aspiration (RPFNA). Ki-67 expression is a possible risk biomarker and is currently being used as a response biomarker in chemoprevention trials. We have previously shown that Ki-67 expression is higher in RPFNA specimens of benign breast cells exhibiting cytologic atypia. It is not known whether there is a correlation between mammographic density and Ki-67 expression in benign breast ductal cells obtained by RPFNA. Methods: 344 women at high risk of developing breast cancer (based on personal or family history) seen at The University of Kansas Medical Center high risk breast clinic, who underwent RPFNA with cytomorphology and Ki-67 assessment, plus a mammogram were included in the study. Mammographic breast density was assessed using the Cumulus program. Categorical variables were analyzed by Chi-square test and continuous variables were analyzed by non-parametric test and linear regression. Results: 40% of women were premenopausal, 7% perimenopausal and 53% were postmenopausal. Median age was 49 years, median 5 year Gail Risk was 2.2%, and median Ki-67 was 1.9%. Median mammographic breast density was 37%. Ki-67 expression increased with cytologic abnormality and number of cells collected, but was unrelated to Gail risk (as observed previously). Breast density was higher in pre-menopausal women (p=0.001), those with lower BMI (p< 0.001), and lower 5-year Gail risk (p=0.012); Breast density showed no correlation with Ki-67 expression or cytomorphology. Conclusion: Given the lack of correlation of mammographic breast density with either cytomorphology or Ki-67 expression in RPFNA specimens, mammographic density and Ki-67 expression should be considered as potentially complementary response biomarkers for breast cancer chemoprevention trials.

No significant financial relationships to disclose.
Aromatase inhibition and very low dose estradiol add-back: A pilot study for novel breast cancer prevention
Authors: Kendall A. , Smith I. , Folkerd E. , Dowsett M.
Published: March 2020

1014

Background: Aromatase inhibitors (AIs) are important agents for the treatment of hormone receptor positive breast cancer in post menopausal women. They are being assessed as chemopreventives of breast cancer in large clinical trials, but there are known issues of tolerability notably affecting bone. It is possible that decreasing, but not abolishing, circulating estradiol levels may reduce breast cancer risk without such side effects. We have undertaken a novel pilot study to establish whether healthy volunteers on an AI could be delivered very low dose estradiol replacement to a predictable serum level of 10–20 pmol/L using a transdermal patch, and whether this would reverse changes in the bone resorption biomarker CTx. Methods: Ten healthy postmenopausal women with normal estradiol levels received letrozole 2.5mg/day for 16 weeks. From weeks 6–16 they were also given a quarter of Estraderm MX25 transdermal patch (6.25 μg of estradiol) replaced, twice weekly. Fasting blood samples were taken at baseline, 6, 10, 12, 14 and 16 weeks for estradiol, FSH, LH, SHBG and CTx. Results: Serum estradiol levels fell to <3pmol/L at 6 weeks consistent with AI therapy. The quarter patch successfully replaced estradiol levels to a median of 12pmol/L although there was marked inter and intrasubject variability. FSH rose significantly with AI therapy with no appreciable change in SHBG or LH. There is encouraging evidence that CTx levels which increased with AI (p=0.076) can be corrected with low dose estradiol replacement although this did not reach statistical significance in this initial group. We continue to collect data on a further 8 subjects already recruited into the study. Conclusions: A quarter of Estraderm MX25 transdermal patch can be used to provide a serum estradiol level of 10–20 pmol/l in postmenopausal women given an AI. Further data will confirm whether this novel approach to chemoprevention can reverse the negative impact of AIs on bone metabolism and be suitable for extending to a full chemoprevention trial.

[Table: see text]

No significant financial relationships to disclose.
Cancer screening trial to evaluate the efficacy of FDG-PET in healthy subjects: 2-year results of the Hamamatsu Medical Imaging Center study
Authors: Nishizawa S. , Inubushi M. , Okada H. , Ozawa F. , Kojima S. , Teramukai S. , Fukushima M.
Published: March 2020

1025

Background: Positron emission tomography with 18F-fluorodeoxyglucose (FDG-PET) is increasingly used for cancer screening in Japan. Although it is not rare that cancers at early stages are incidentally found on FDG-PET in usual clinical practice, it is still unknown whether FDG-PET is an effective tool for early detection of cancer and decrease of cancer death. To evaluate the efficacy of FDG-PET in cancer screening, we started a prospective study 2 years ago. Methods: From Aug 26 2003 to July 22 2004, 1197 healthy volunteers aged 35 years or older (46.7±7.7y.o.) participated in our cancer screening trial. They are supposed to undergo screening tests annually for 5 years and will be followed for years. The screening tests included whole-body FDG-PET, chest and abdominal CT, brain and pelvic MRI, tumor markers (CEA, CA19–9, SCC, PSA, CA125), and fecal occult blood test. From July 26 2004 to July 22 2005, 1158 subjects of the 1197 underwent the second screening a year after the first one. We made contact with 39 other subjects who skipped the second screening to check the occurrence of cancers. Results: In the 2-year period, primary cancers occurred in 19 subjects. Fifteen cancers (6 thyroid, 3 breast, 2 prostate, 3 lung, 1 ectopic thymus in thyroid grand) were detected on the first year and one (prostate) on the second year by the screening. Ten of the 16 were at stage I. Three subjects with negative findings at the screening developed gastric lymphoma, gastric cancer and malignant lymphoma of the kidney, respectively, within a year after the screening. FDG-PET was positive in 10 of 15 cancers (6 thyroid, 2 breast, 1 prostate and 1 ectopic thymus) on the first year. No PET positive cancer was found on the second year. Sensitivity of the screening tests as a whole and FDG-PET alone for the first year was 78.9% and 52.6%, respectively. Positive and negative predictive values and specificity of FDG-PET were 11.0% (10/91), 99.2% (1097/1106), and 93.1% (1097/1178), respectively. Conclusions: Cancer screening including FDG-PET showed reasonable accuracy and could detect a wide variety of cancers at early stages. This prospective study will clarify the details of efficacy and suitable interval of FDG-PET in cancer screening.

No significant financial relationships to disclose.
Preference and compliance of stool occult blood testing in a rural cancer-screening program
Authors: Waheed S. , Linthacum T. A.
Published: March 2020

1030

Background: Colon cancer is the third most common cancer in men and women. Screening for colon cancer reduces the incidence of colon cancer; decreases mortality associated with colon cancer and should be an integral part of all comprehensive cancer screening programs. The impact of patient preference and compliance when given the option between guaiac-based fecal occult smear method and immunochemical assay testing was reviewed at a rural cancer-screening clinic. Methods: All cancer screening participants over age 50 were educated about colon cancer prevention and screening. The immunochemical assay test and the guaiac-based fecal occult blood test directions and costs were thoroughly explained. A sample test packet of the chosen method was shown to the participant for further explanation on the directions. Data was collected for over one year with documentation of type of preference of test taken and the return rates of those tests. Results: A total of 2170 immunochemical assay or guaiac-based fecal occult blood tests were offered to participants over age 50. The total percentage of tests taken was 39% (852 participants). 81% (688 participants) of the participants chose the immunochemical test over the guaiac-based fecal occult blood test 18% (155 participants). The return rate continued in favor of the immunochemical test with 86% (380 participants) return rate as compared with only 14% return rate for guaiac-based fecal occult blood test (60 participants). The total return rate for both test combined was 51%. Conclusion: This study revealed a surprising lack of acceptance of colon caner screening by means of stool occult blood testing despite vast efforts at thorough explanations and education. When occult blood testing was accepted immunochemical assay testing was the preferred method. Further research should focus on patient barriers in accepting colon cancer testing.

No significant financial relationships to disclose.
Evaluation of FDG PET in early axillary lymph node response to neoadjuvant chemotherapy in locally advanced breast cancer patients
Authors: Rousseau C. , Bodet-Milin C. , Bennouna J. , Ferrer L. , Campion L. , Bridji B. , Sagan C. , Ricaud M. , Resche I. , Kraeber-Bodere F. , Campone M.
Published: March 2020

10500

Background: Regional axillary lymph node status has remained the single most independent variable to predict prognosis both in terms of disease recurrence and survival. This study aimed to prospectively assess sequential FDG PET findings as early predictors of axillary lymph node response to neoadjuvant chemotherapy in locally advanced breast cancer patients. Methods: A total of 255 attenuation-corrected scans were obtained from 51 patients. Images were acquired with a PET/CT scanner (GE Discovery LS) following administration of FDG (5 MBq/kg) at baseline, and after the first, second, third, and sixth course of chemotherapy. Clinical examination and US were used to assess the size of axillary lymph nodes. PET data were interpreted using standardized uptake values (SUV) corrected or not for partial volume effects. Changes in the SUV were considered as surgical/histopathological responses. Results: This study focused on axillary lymph node staging, therefore, primary breast tumor data were not considered in the analysis. The sensitivity, specificity, and accuracy of axillary node staging was higher with PET (75%, 87%, 80%) than with clinical examination and US (50%, 78%, 63%, and 50%, 83%, and 65%, respectively), and even the more so when PET images were corrected for partial volume effects (86%, 83%, 84%). Significant differences between absolute FDG uptake values in responders and non-responders were observed at baseline (p < 0.05). While FDG uptake did not vary much in non-reponders, as confirmed by histopathology, it markedly decreased to baseline levels in responders (p < 10−5). Fifty percent of baseline SUV was considered the best cut-off value to distinguish responders from non-responders. The sensitivity, specificity, negative predictive value, and accuracy of FDG PET after one course of chemotherapy were respectively, 96%, 75%, 95%, and 84%. Tumor response assessment by clinical examination or US was never statistically significant whatever the cut-off. Conclusions: The pathological status of regional axillary lymph nodes in locally advanced breast cancer patients can be accurately predicted after one course of neoadjuvant chemotherapy based on FDG PET images.

No significant financial relationships to disclose.
Multicenter phase II study of a cremophor-free polymeric micelle-formulated paclitaxel in patients (pts) with metastatic breast cancer (MBC)
Authors: Lee K. S. , Chung H. C. , Im S. A. , Park Y. H. , Kim S. B. , Kim C. S. , Ro J.
Published: March 2020

10520

Background: The results of a phase I trial of a novel taxane, a cremophor-free polymeric micelle-formulated paclitaxel (Genexol-PM), were previously reported (Clin Cancer Res 10:3708, 2004). The dose limiting toxicities were neuropathy, myalgia, and neutropenia. Notably, acute hypersensitivity reaction was not observed. Methods: This was a single arm phase II study performed at 6 major hospitals in Korea to evaluate the efficacy and safety of Genexol-PM as a 1st- or 2nd-line therapy for MBC. Genexol-PM with no premedication was given at 300 mg/m2 i.v. over 3 hr on day 1 every 3 weeks until disease progression or intolerability. Two-stage group sequential design was used. If more than 14 responses were attained in the first 30 pts, early termination was possible. Results: Forty-three pts with MBC were enrolled from 7/2004 to 12/2004. Median follow-up duration was 7.0 mo. Thirty-seven of 41 pts excluding 2 pts who did not receive Genexol-PM received Genexol-PM as a first-line therapy. Nineteen pts received prior anthracycline containing regimen in adjuvant (15) and in metastatic (4) settings. No pts were anthracycline-resistant. Median age was 48 years (range; 30 to 69 years) with ECOG PS 0 or 1. A total of 331 cycles were administered, with median of 8 per patient (range; 1 to 16). Of 39 pts evaluable for efficacy, there were 5 CRs /19 PRs with response rate of 61.5%, 13 SDs and 2 PDs. Median response duration was not reached (range; 1.9+ to 10.0+ mo). Median TTP and OS were 9.8 and 11.2 mo, respectively. G3 non-hematologic toxicities included peripheral sensory neuropathy (51.2%) and myalgia (2.4%). Hematologic toxicities were G3/G4 neutropenia (51.2%, 17.1%, respectively) and G1/2 thrombocytopenia (22.0%). Seventeen pts (41.5%) required dose reductions due to G3 neuropathy. No febrile neutropenia was observed. Eight pts developed hypersensitivity reactions with G3 in 2 pts. Mean dose intensity (DI) was 87.4 mg/m2/wk (range; 27.7 to 116.7) at 87.4% of planned DI. Conclusions: Genexol-PM at 300 mg/m2 appears a promising agent with high response rate for anthracycline sensitive MBC. Further trials at a different dose schedule or duration of delivery, or in combination with other drugs are warranted. Supported by Samyang Co, Korea.

No significant financial relationships to disclose.
Does adjuvant chemotherapy (CT) exacerbate cognitive impairment in elderly breast cancer (BC) patients? Results of a prospective, longitudinal study
Authors: Rodin M. B. , Wallace J. A. , Lacy M. , Kuball K. , Pykkonen B. , Fleming G.
Published: March 2020

10530

Background: Over the last few years, several retrospective studies documented cognitive impairment in post-chemotherapy BC patients. This decline was hypothesized to be related to CT (“chemo-brain”). More recently, a prospective study (Wefel et al. 2005) documented significant cognitive impairment at baseline, which may impact response to CT, along with treatment related effects. While all previous studies focused on the cognitive response of young BC women, older patients may be much more susceptible to any CT related neurotoxicity. Thus, the current study investigated the incidence and course of cognitive impairment in older, post-menopausal BC patients undergoing CT. Methods: 19 postmenopausal women (≥50 yrs) with no prior chemo or hormonal cancer treatment, and baseline MMSE >23 completed a comprehensive neurocognitive battery of tests along with anxiety and depression measures, prior to treatment. At 6 months, 12 patients returned for post-chemotherapy evaluation, with 8 evaluated at 2 years. Ten DCIS patients were also evaluated at baseline and 6 months. Results: Mean age at baseline was 67 yrs with 14 years of education. Prior to treatment, 37% displayed significant cognitive impairment not accounted for by other variables. At six months, within and between group analyses revealed only improved fine motor speed (Finger tapping, p.01) related to practice effects, but no decline in cognitive functions. There were no significant differences across measures between BC and DCIS groups. However, RCI revealed 3 of 12 BC patients improved on memory tasks. Two year longitudinal analyses again revealed improved motor speed and possible cognitive flexibility (TMT-A, p.02), along with decreased anxiety (STAI-S, p = .03). Conclusion: This small study does not support a finding of treatment-associated cognitive impairment among elderly women receiving standard CT for breast cancer. However, like Wefel’s sample, we documented significant cognitive impairment prior to initiation of treatment. Studies addressing the etiologic mechanism related to this cognitive dysfunction are warranted. Limitations of the current study will be discussed.

No significant financial relationships to disclose.
Preoperative prediction of the axillary lymph nodes status in patients with breast cancer
Authors: Angelidou E. , Sotiropoulou G. , Poulianou E. , Politi E. , Koutselini H.
Published: March 2020

10547

Background: We developed a preoperative score-system (S) and evaluated prospectively its predictive value for the axillary(a) status of patients (p) with breast cancer. Our aim was to select preoperatively (p) with negative axilla, who could possibly avoid the standard (a) surgery. (S) uses preoperative clinical, epidemiological and immunocytological data, obtained from the FNA-smears of (p)‘ tumors, and attempts to guide the choice of (a) treatment, as an alternative to the method of sentinel lymph node. Methods: (S) is calculated by adding the preoperative values of clinical tumor size, (p) age, nuclear grade (NG), type of the cancer cells and the immunocytochemical positiveness of the biomarkers p53, HER2 and MIB1. Values range from 1–4 for size (1–10, 11–15, 16–20, 20–30 mm), 1 to 4 for age (70 and over, 51–69, 41–50, 40 years or less), 1–3 for NG1–3, 1–2 for type of cancer cells (lobular, ductal) respectively and 0–3 for the expression (1 point for every positive biomarker) or absence (0) of p53, HER2 and MIB1 in the FNA of the primary tumors of the (p). (S) ranges from 4 to 16.We applied (S) to 224 (p), with clinically negative axilla. These (p) underwent modified radical mastectomy or lumpectomy and standard (a) dissection level I and II. The number of the infiltrated nodes was identified in each case. Results: (S) of 4 - 8 (57 patients, group 1) identify (p) with free nodes ( node positive rate 0%). (S) of 9 and 10 (67 patients, group 2) carry an average node positive rate of 65,67%, of which 31,34% involves the invasion of 1 node, 23,88% of 2–3 and 10,44% of 4 or more nodes (P < 0.001, group 1 versus group 2). (S) of 11 and more (100 patients, group 3) identify (p) with an average node positive rate of 83%, of which 55% involves the invasion of 4 or more nodes (P < 0.001, group 3 versus group 1). (S) allows the separation of (p) into two (a) management groups. Group 1 are those (p), who possibly have free lymph nodes and therefore may need no (a) surgery at all, whereas group 2 and 3 may be considered for standard (a) dissection, because they present with increased possibility infiltrated nodes. Conclusion: (S) was studied to aid the selection of (p) towards reasonable (a) treatment choices for the benefit of (p). (S) might serve as a guideline in the clinical practice to reduce the postoperative morbidity of the breast cancer (p).

No significant financial relationships to disclose.