bluuebunny/crossref_metadata_2025_split huggingface dataset:
MicroRNA-based assay for differential diagnosis of squamous from non-squamous non-small cell lung carcinoma
Authors: Lebanony D. , Benjamin H. , Gilad S. , Ashkenazi K. , Nonaka D. , Feinmesser M. , Rosenfeld N. , Chajut A. , Cohen D. , Aharonov R. 
Published: August 2020

11069 Background: Recent advances in biologically directed therapies for non-small cell lung carcinoma (NSCLC) emphasize the need for more accurate sub-classification of NSCLC, as treatment may be dictated by histologic subtype. In particular, squamous histology can be a counter-indication for treatment by VEGF inhibitors. MicroRNAs are highly tissue-specific biomarkers with potential clinical applicability for defining cancer type and origin. MicroRNAs are well preserved in formalin fixed tissue, making them ideal candidates for molecular markers for use in routinely processed material. Here we report on the development and performance of a microRNA-based assay for the differential diagnosis of squamous from non-squamous NSCLC. Methods: We developed protocols for extraction of high-quality RNA that retain the microRNA fraction from FFPE archival tissue samples. MicroRNA microarrays were used to profile more than a hundred NSCLC samples. Specific microRNA qRT-PCR was used to validate results, and to develop a diagnostic assay. Results: We identified a microRNA biomarker that is strongly overexpressed in squamous cell NSCLC. A diagnostic assay (miRview squamous) was developed, that utilizes qRT-PCR measurement of this microRNA, normalized by an additional microRNA and a small nuclear RNA. This assay was validated on a blinded test set of 64 tumor samples, and had sensitivity of 97% and specificity of 91%. More than ¾ of the samples were classified with high confidence, and these classifications were accurate in 96% of the cases. Conclusions: MicroRNAs are becoming an important tool for classification of cancers. A diagnostic assay based on the specificity of a single microRNA accurately identifies squamous from non-squamous NSCLC. This assay provides an important new tool for the classification of NSCLC. [Table: see text]
Role of Galectin-3 in the activation of p-ERK and in resistance to radiotherapy in glioblastoma through the activation of p-ERK independent of Ras activity
Authors: El-Jawahri A. R. , Lautenschlaeger T. , Patel D. , Chakravarti A. 
Published: August 2020

11073 Background: There has been a growing interest in investigating the molecular mechanisms leading to treatment resistance in glioblastoma (GBM). The MAPK pathway has been shown to be involved in mediating treatment resistance. Galectin-3 (Gal3) is commonly overexpressed in astrocytic tumors of the brain and has been associated with adverse clinical outcomes. We investigated the role and mechanisms of Gal3 in mediating treatment resistance in GBM through modulating the MAPK pathway. Methods: We investigated Gal3 expression in GBM cell lines (U87, LN18, LN229). We created stable Gal3 knockdown cell lines (Gal3KD), ERK-knockdown (lentiviral shRNA) and constitutively active MEK(MEK2DD plasmid obtained from Dr. Brugge) cell lines. We then evaluated the radiosensitivity, apoptosis, and cell viability of control cell lines treated by non-targeting (NT) vectors as compared to Gal3KD, ERKKD and MEK2DD using clonogenic survival, Annexin V/PI apoptosis, COMET, and cell cycle assays. Signaling cascades were evaluated using Western blot and a GTP-ras pull down assay. Gal3 was also transiently knocked down in MEK2DD and NT cell lines and signaling cascades were analyzed using western blotting. Gal3 binding interactions were identified using co-immunoprecipitation. Results: Gal3 expression was induced in all GBM cell lines following irradiation. Gal3KD showed increased radiosensitivity similar to ERKKD as compared to controls with decreased clonogenic survival, increased DNA damage, increased apoptosis, and increased cell cycle arrest in G1 and G2, while MEK2DD showed enhanced radioresistance. Gal3KD showed reduced p-ERK at baseline followed by a minimal induction of p-ERK with irradiation as compared to controls which have a high level of p-ERK. Overall p-Akt, ERK and Ras-GTP levels were equivalent in Gal3KD and controls. When Gal3 was transiently knocked down in MEK2DD cell line, p-ERK was drastically reduced similar to controls. Gal3 co-immunoprecipitated with ERK. Conclusions: Our results suggest a possible direct novel interaction between Gal3 and ERK in GBMs that is essential for the expression of p-ERK and mediating treatment resistance in GBMs. No significant financial relationships to disclose.
Combining multigene profiling of molecular subtypes with the 70-gene profile for classification of breast cancer
Authors: Stork-Sloots L. , Krijgsman O. , Roepman P. , de Snoo F. A. , Bender R. A. , Glas A. M. 
Published: August 2020

11083 Background: Classification of breast cancers into molecular subtypes may be important for accurate selection of therapy for patients. Herein we report a multigene profile for classification of breast cancer into molecular subtypes. The profile separates tumors into hormone receptor (HR)+/luminal-like, HER2+/ERBB2-like, and triple negative/basal-like subclasses. Methods: A multi-gene profile was developed based on a series of 200 tumor samples of known ER, PR and HER2 receptor status (concordant IHC and gene expression result) hybridized on 44k microarrays. The profile classifies 96% concordant to the molecular subtypes named luminal-, ERBB2- or basal-type as published by Perou et al (Perou et al, Nature, 2000; Fan et al, NEJM, 2005). The profile was validated using 469 independent samples as well as on two publically available gene expression datasets (n=251 and n=159). Results: The profile classified 66% (712) as luminal-like, 18% (194) ERBB2-like, and 16% (173) as basal-like. As compared to single-marker readout for the presence of ER, PR and HER2, 13% of the samples that were scored positive for presence of ER/PR did not express a luminal-like gene profile. Samples with a ERBB2-like or basal-like gene profile showed equally poor 5-year survival rates of ∼65%. However, the ERBB2-like subset of MammaPrint low risk patients (15%) showed an 89% (95%CI, 71–100) survival rate without trastuzumab treatment. When the luminal-like subtype was separated into “high-” and “low-risk” by MammaPrint the survival rate was 56% (95%CI, 46–68) for high-risk luminal-like samples and 94% (95%CI, 90–99) for low-risk samples. Conclusions: The developed multigene profile can classify breast tumors into luminal-, ERBB2- and basal-like subgroups. By combining this molecular subtyping with MammaPrint risk-classification specific groups of patients can be recognized that that are at high risk of recurrence. The low risk patients within the luminal- and ERBB2-like subclasses have a very low risk of recurrence. Implementation of this knowledge can improve the clinical management of breast cancer patients. [Table: see text]
Adjuvant UK: Validation of a UK prognostic model for early breast cancer
Authors: Wishart G. , Greenberg D. , Rashbass J. , Ravdin P. , Lawrence G. , Caldas C. , Pharoah P. 
Published: August 2020

11086 Background: To develop and validate a prognostication model to predict overall survival for women treated for early breast cancer in the UK based on cancer registry data. The main advantages are that unlike SEER, this dataset includes accurate information on mode of detection as well as systemic therapy. Methods: Using the Eastern Cancer Registration & Information Centre (ECRIC) dataset, information was collated for 5,818 women diagnosed with invasive breast cancer in East Anglia from 1999–2003. All patients underwent surgery, had records of pathological staging (tumour size, grade, lymph node status, ER status) and systemic treatment (chemotherapy, hormone therapy, both), mode of detection (screen-detected, symptomatic) and were followed up to December 2007. A model was derived from these data using Cox proportional hazards and this was subsequently validated in an external dataset of 5,468 patients from the West Midlands Cancer Intelligence Unit (WMCIU). Validation was performed by comparing (a) observed and predicted mortality - overall and by risk group decile - and (b) receiver operating characteristic (ROC) curves in both ECRIC & WMCIU datasets. Results: ECRIC dataset: Observed/predicted deaths were: 2 years (262/267), 5 years (868/867), 8 years (1108/1093). Area under ROC curve was 0.800WMCIU dataset: Observed/predicted deaths were: 2 years (317/298), 5 years (862/919), 8 years (955/999). Area under ROC curve was 0.783. The model and validation worked equally well at all time points (2, 5, 8 years) and in all risk group deciles. In the validation dataset the observed and predicted 8-year OS was within 1% (17.5% versus 18.3%). There was no significant statistical difference between the ROC curves (X2 = 2.35, P=.12) for prediction at 8 years in ECRIC and WMCIU data. Conclusions: We have developed a prognostication model for early breast cancer based on data from a UK cancer registry and validated the model using data from another UK registry. The model was well calibrated and provides a high degree of discrimination. This model will underpin the first web-based prognostication and treatment benefit tool for early breast cancer in the UK (Adjuvant UK). No significant financial relationships to disclose.
Prognostic value of the genomic grade index (GGi) as compared to centrally measured Ki-67 IHC and mitotic index in early breast cancer patients
Authors: Bertucci F. , Le Doussal J. M. , Birnbaum D. , Tagett R. , Martinec A. , Hermitte F. , Marisa L. , Martin A. L. , Geneve J. , Roché H. , Penault-Llorca F. 
Published: August 2020

11094 Background: Genomic grading has been proposed to improve tumor grading. The genomic grade index (GGi) is a 97-gene continuous measure which resolves 80% of histological grade 2 (HG 2) tumors into HG 1 and HG 3 risk categories. GGi has higher prognostic value than HG in patients treated with and without systemic adjuvant endocrine and chemotherapy. A key issue is whether the GGi adds prognostic information to centrally-determined mitotic index and Ki-67 IHC. Methods: The control arm of the PACS 01 trial included 996 women with node-positive (N+) early breast cancer treated with six cycles of fluorouracil, epirubicin, and cyclophosphamide (FEC) and tamoxifen as required. 128 genomic profiles could be obtained from available frozen tumor samples using Affymetrix U133 Plus 2.0 gene chips through the “Carte d'Identite des Tumeurs” program of the French Ligue Nationale contre le Cancer. The Genomic Grade index (GGi) was computed using Ipsogen MapQuant Dx(R). Central Elston-Ellis grade, mitotic index (mitosis / mm2), and Ki-67 IHC (% positive cells) were available for 125 patients. The GGi and histological parameters were correlated to the 5-year metastasis status (MFS-5) by ROC analysis and to the metastasis hazard (follow-up of 6.2 ± years) by Cox regression. Results: In ER+ patients (n=93), the GGi was the only significant correlate to metastasis hazard in multivariate Cox regression with histological parameters (HR = 3.5 [1.7–7.5], p<0.001). It was the best predictor of MFS-5 (ROC AUC = 0.83, p=1E-6) when compared to histological parameters (ROC AUC = 0.71, 0.72 and 0.66 resp. p=0.003 to 0.03). In HG 2 subgroup (n=43), the GGi was the only significant predictor of MFS-5 (ROC AUC = 0.81, p=0.016). Conclusions: In our sample of N+ ER+ breast cancer patients, the GGi improved prognostication compared to centrally-measured mitotic index and Ki-67 IHC used alone and in combination. Moreover, the GGi was the only prognostic factor in histological grade 2 patients. [Table: see text]
Thymidylate synthase polymorphisms and immunohistochemistry in non-small cell lung cancer
Authors: Stoehlmacher J. , Goekkurt E. , Hoeffken G. , Zinsky R. , Lynch F. , Buettner H. , Scheil-Bertram S. , Schirren J. , Ehninger G. , Fisseler-Eckhoff A. 
Published: August 2020

11101 Background: Recently, pemetrexed has been introduced into the treatment of non-small cell lung cancer (NSCLC). There's evidence that pemetrexed appears to be more efficient in non squamous NSCLC including adenocarcinoma (AC) and large cell carcinoma (LCC). TS represents a major target enzyme of pemetrexed. We evaluated the expression of TS among different histological types of NSCLC and its association with functional genetic polymorphisms of the TS gene. Methods: Archived tumor samples from 312 individuals with NSCLC were analyzed. Immunohistochemistry (IHC) was performed using a mouse monoclonal antibody to TS. TS stained sections were scored for cytoplasmic and nuclear localization. For each compartment, the tumor was evaluated for the estimated percentage of positive cells with the greatest intensity within the tumor. Intensity was scored on a scale of 0–4. A score of 3–4 was classified as high expression. Genotyping of TS-VNTR including the G/C SNP and TS1394del6 was performed by PCR based RFLP techniques. Genotypes supposed to be associated with low expression were grouped (group A: 2R/2R or 2R/3RC or 3RC/3RC + -6/-6) and compared to genotype groups associated with high expression (group B: 2R/3RG or 3RC/3RG or 3RG/3RG + -6/+6 or +6/+6). Results: IHC and polymorphisms could be performed successfully in 312 (100%) and 287 (92%), respectively. Distribution of histology was as follows: 50% AC, 42% squamous cell carcinoma (SCC), 3% LCC and 5% mixed or other histological subtypes. Group B genotypes were significantly more present in SCC than in AC (49% vs 23%, p=.002). Tumors with group B genotypes were more likely to display high nuclear staining intensity than group A tumors (30% vs. 17%, p=.077). A similar but not significant trend was seen for cytoplasmic staining intensity. There was no significant difference between histological subtypes of NSCLC with respect to TS protein expression. AC were more common than SCC to show a G1 differentiated tumors (8% vs 1%, p<.001). No difference between genotype groups were observed with respect to grading. Conclusions: TS polymorphisms are significantly associated with histology subtypes in NSCLC. These results represent a potential explanation for efficacy differences of pemetrexed in NSCLC and warrants further investigations. [Table: see text]
Evaluation of aldehyde dehydrogenase 1 (ALDH1) as a marker of non-small cell lung cancer (NSCLC) stem cells (SCs) and correlation with prognosis
Authors: Zhou R. , Liu Q. , Todd N. W. , Deepak J. , Liu Z. , Stass S. A. , Edelman M. J. , Katz R. L. , Jiang F. 
Published: August 2020

11105 Background: ALDH1 is a cytosolic enzyme responsible for oxidizing intracellular aldehydes, and conversion of retinol to retinoic acid in SCs. ALDH1 has been previously demonstrated to be a marker for SCs in breast cancer (Ginestier C, et al., Cell Stem Cell. 2007 Nov; 1(5):555–67). The Aldefluor assay is based on conversion of a synthetic substrate BAAA, when passively infused into cytosol, to brightly fluorescent BAA by ALDH1. Combined with fluorescence-activated cell sorting (FACS), it has been used successfully in hematopoietic and breast cancer SCs isolation. ALDH1 is also found in NSCLC. We hypothesized that ALDH1 would be a marker for NSCLC SCs and a potential prognostic marker. Methods: NSCLC SCs were isolated from human NSCLC cell lines using the Aldefluor assay and FACS. ALDH1-positive cells were analyzed extensively for SC characteristics. ALDH1 expression in 303 NSCLC biopsy specimens from three independent cohorts of NSCLC patients was then analyzed by immunohistochemisty (IHC) using an ALDH1 antibody (Santa Cruz Biotechnology) and commercially available negative controls. Results: Isolated cancer cells with high ALDH1 activity displayed cancer SCs features, including capacities for proliferation, self-renewal, differentiation; resistance to chemotherapeutic agents including cisplatin, vinorelbine, gemcitabine and docetaxel; expression of the SC surface marker CD133, and high invasiveness. ALDH1-positive cells generated tumors in vivo recapitulating heterogeneity of parental cells. Xenograft tumors derived from ALDH1-positive cells (103 each) were 36±2.9 mm3 on average in size, with some of the cells having lost ALDH1 expression. Xenograft tumors derived from ALDH1-negative cells (105 each) were 4 mm3 on average in size, without ALDH1 reactivation. Statistical analysis of quantitative IHC and clinical data showed ALDH1 expression was correlated with higher stage and grade of NSCLC (p = 0.02). Expression of ALDH1 in stage I NSCLC patients was linked to decreased 5 year cancer-specific survival (62% vs. 96%, p = 0.006) and overall survival (32% vs. 72%, p = 0.009). Conclusions: 1. ALDH1 is a NSCLC SC-associated tumor marker. 2. ALDH1 expression is a negative prognostic marker in early stage NSCLC. No significant financial relationships to disclose.
Topographical analysis of telomere length and correlation with genomic instability in whole mount prostatectomies
Authors: Joshua A. M. , Marrano P. , Evans A. , Van der Kwast T. , Zielenska M. , Squire J. 
Published: August 2020

11107 Background: Many critical events in prostatic carcinogenesis appear to relate to the emergence of chromosomal instability and acquisition of genomic rearrangements. Characteristic abnormalities such as 8p loss, 8q gain, trisomy 7, PTEN microdeletions and TMPRSS2-ERG gene fusions appear to mediate mechanisms to increase neoplastic transformation in prostate cancer. Current evidence suggests that telomere dysfunction is a likely causative factor for some of these abnormalities on the basis of its relationship to mechanisms such as the break-fusion-bridge cycle that can lead to the onset of chromosomal instability. Methods: In this study, we correlated telomere length in various prostatic histologies by quantitative FISH with genomic markers of chromosomal instability by standard FISH and immunohistochemical measures of proliferation in 3 whole mount prostatectomies. Results: After analysing approximately 25,000 cells, we found that telomere shortening was correlated with an increase in the number of cells with abnormalities on chromosome 8, such as an increase in the average number of c-myc signals (r∼0.35, p∼0.02). However, there were no significant correlations with abnormalities such as trisomy 7 or abnormalities of the PTEN locus in any sample. Additional findings included; associations found with the probability of C-MYC aberrations in stroma with greater proximity to cancer (<1,000 um), a correlation between telomere length in a number of prostatic histologies (normal, atrophy, HPIN and cancer) with the adjacent stroma, and a lack of correlation between the Ki67 index of various histologies and their telomere length - all suggesting the importance of microenvironmental effects on telomere maintenance in the prostate. Finally, we also report significant telomere shortening in BPH in 2 cases, a phenomenon that has not been noted previously. Conclusions: This is the first study to directly link a mechanism of chromosomal instability with specific chromosomal abnormalities in prostatic carcinogenesis and also suggests that the microenvironmental milieu is of critical importance in the evolution of in vivo telomere homeostasis. No significant financial relationships to disclose.
Use of the ErbB2/CEP17 ratio to predict prognosis and response to trastuzumab-based therapy in the metastatic breast cancer setting
Authors: Fuchs E. , Köstler W. , Horvath R. , Hudelist G. , Kubista E. , Attems J. , Zielinski C. , Singer C. F. 
Published: August 2020

11110 Background: Despite patient selection based upon detection of Her-2/neu overexpression by immunohistochemistry (IHC) and/or presence of amplification of the Her-2/neu-encoding erbB2 oncogene measured by FISH, response to trastuzumab-based therapy is only achieved in a subset of patients with Her-2/neu overexpressing breast carcinomas. Exact quantification of erbB2 copy-number relative to chromosome 17 (CEP17) (ErbB2/CEP17 ratio “R”) probably adds further important predictive information. Methods: Clinical data of 137 patients receiving trastuzumab based treatment for Her-2/neu overexpressing (IHC) metastatic breast cancer were analysed. ErbB2/CEP17 ratio (R) was determined by quantitative FISH analysis in original tumor tissue using Vysis PathVysion DNA-based FISH technology. Results: ErbB2/CEP17 (R) provided additional predictive value for progression free survival (PFS) and time to first metastasis (TTM), but not for overall survival (OS) (all from start of trastuzumab containing treatment). The following cutoffs of Her-2/neu were identified: group A: 0–2.2 R (TTM: 49.8; OS: 6.7; PFS: 6.2); group B: 2.2- 6 R (TTM: 26.2; OS: 5.3; PFS: 9.3); group C: >6 CN (TTM: 20.1; OS: 3.9; PFS: 13.7) Kaplan-Maier analysis showed significant longer TTM for group A (p<0.01 vs. B/C), significant longer PFS for group C (p<0.01 vs. A/B). Significant differences in complete response (B/C: 16.9% vs C:44.4%), partial response (B/C: 20.2% vs. C: 33.3%) and progressive disease (B/C: 27% vs. 11.1%) were noted. No significant difference in overall survival between the groups was seen. Conclusions: ErbB/CEP17 R provides important prognostic information and, in metastatic patients, allows one to better predict response to trastuzumab-based treatment than the widely used binary classification of ErbB2 amplification that is based on a cut-off at a copy number of >2.2. No significant financial relationships to disclose.
The prognostic factors for the recurred breast cancer patients with isolated, limited number of lung metastasis, and the implication of pulmonary metastasectomy
Authors: Yhim H. , Han S. , Oh D. , Han W. , Im S. , Kim T. , Kim Y. , Noh D. , Ha S. , Bang Y. 
Published: August 2020

1112 Background: In recurred breast cancer (BC) with isolated lung metastasis, the role of pulmonary metastasectomy (PM) is not established. Methods: We consecutively enrolled recurred BC patients (pts) with isolated lung metastasis from 1997 to 2007 in Seoul National University Hospital. Among 140 recurred pts with isolated lung metastasis, 45 had metastatic lesions less than 4. We analyzed the clinical outcomes of these 45 pts. Results: The median age was 49.0 (range 32–67) years. The median disease-free interval (DFI, Time from initial BC diagnosis to recur) was 26.1 (range 4.2–168.0) months, and the pts with DFI of less than 24 months were 20 (44%). The pts who had biologic subtypes (BS) of hormone receptor (HR)(+), HER-2(+), triple negative (TN) were 16 (36%), 14 (31%,) and 15 (33%), respectively. Fifteen (33%) had PM then followed by systemic treatment (PM group), and 30 received systemic treatment without PM (no PM group). There was no difference in terms of clinical factors such as DFI and BS between two groups. The median progression-free survival (PFS) and overall survival (OS) were 13.0 months (95% CI, 10.05–15.95) and 41.7 months (95% CI, 23.82–59.58), respectively. In univariate analysis, PFS was significantly longer in pts with PM (p < 0.001), DFI of more than 24 months (p < 0.001), and was marginally significant according to BS (p = 0.084). In multivariate analysis, DFI of less than 24 months (hazard ratio [H], 4.53; 95% CI, 1.72–11.90), no PM (H, 9.52; 95% CI, 3.34–27.18) and BS such as HER-2(+) (H, 3.00; 95% CI, 1.04–8.64) and TN (H, 3.92; 95% CI, 1.32–11.59) were independent prognostic factors for shorter PFS. In univariate analysis, OS was significantly longer in pts with PM (p = 0.011), DFI of more than 24 months (p < 0.001), HR(+) BS (p = 0.009). In multivariate analysis, DFI of less than 24 months (H, 7.67; 95% CI, 2.00–29.43) and BS such as HER-2(+) (H, 3.70; 95% CI, 1.03–13.24) and TN (H, 3.27; 95% CI, 1.02–10.44) were independent prognostic factors for shorter OS. Conclusions: In recurred BC with isolated, limited number (less than 4) of lung metastasis, DFI and BS were prognostic factors for PFS and OS. PM also prolonged PFS, and, for OS, more follow-up is needed. Further prospective studies to define the role of PM are warranted. No significant financial relationships to disclose.