bluuebunny/crossref_metadata_2025_split huggingface dataset:
Retrospective immunohistochemical study of VDR, RXR, Ki-67, and Bcl-2 expression in primary and metastatic osteosarcoma
Authors: Tawfik O. , Garimella R. , Tancabelic J. , Keighley J. , Pinson D. , Khan Q. , Templeton K. 
Published: August 2020

e21516 Background: Osteosarcoma (OS) is a highly malignant tumor with a peak incidence in adolescents and young adults. Patient's prognosis is limited to clinical parameters whereas molecular markers of tumor aggression are yet to be identified. Vitamin D (Vit D) has been postulated as a novel therapeutic option for a variety of malignancies such as breast and prostatic carcinomas. The biologic effect of vitamins is mainly mediated through receptors. Vit D receptors (VDRs) are members of the steroid hormone superfamily and are ubiquitous in a various tissues including bone. Upon activation by its ligand, VDR combines with retinoid receptor (RXR) forming a heterodimer initiating a cascade of cellular events. How Vit D induces its antineoplastic activity in OS remains to be elucidated. Methods: Immunohistochemical analysis for VDR, RXR, Ki-67, and bcl-2 was performed on 87 OS specimens to evaluate differentiation, proliferation and apoptosis. Clinical data including site of the primary tumor, presence or absence of metastasis, therapeutic regimens, and outcome were recorded. Results: The mean age of OS patients is 25.7 years (range 7–68). Fifty-four patients were <30 years. Thirty-five OSs were conventional, 17 chondroblastic, 6 giant cell, 6 fibroblastic, 6 mixed, 3 telangiectatic, and 2 epithelioid variants. Twelve of the 87 samples were metastatic. All metastatic OSs were high grade, located in the lungs with a mean age of 25 years (range 7–52). The majority of the OSs expressed VDR (74/87, 85%) and RXR (72/78, 92%). Proliferative activity varied between tumors types with highest Ki-67 percentage noticed in conventional variant (29%), followed by chondroblastic variant (16%). Metastatic tumors consistently had the highest proliferative activity as compared to primary tumors (35%). None of the tumors studied was immunoreactive for bcl-2. Conclusions: Our results demonstrate that VDR and RXR are expressed on most OSs. A significant relationship exists between tumor histologic types and proliferative activity. Metastatic OSs appears to have a significantly higher level of proliferative activity as compared to primary tumors. These results would establish a foundation for elucidating mechanisms by which Vit D induces its antineoplastic activity in OS. No significant financial relationships to disclose.
Clinical and pathological study on 88 cases of very low-risk gastrointestinal stromal tumors
Authors: Hou Y. , Zhou Y. , Lu S. , Tan Y. 
Published: August 2020

e21521 Tumor size <2 cm and mitotic figures <5 per 50 high-power field are considered as very-low-risk GIST according to the NIH consensus criteria, but this two indices can not guarantee a benign clinical course with prolonged follow-up. We investigate whether there are morphological parameters associating with the biological behavior in the very-low-risk GISTs. 88 tumors met the very-low-risk criteria were enrolled for this study. Among them, 80 patients were followed up ranging from 1 to 16.3 years, 5 patients developed relapse. Microscopically, definite infiltrative growth pattern such as muscularis propria, muscularis mucosa or nerve infiltration were identified in 4 of 5 patients, 3 of the 4 patients experienced multiple recurrences. Infiltrative growth characters became more extensive in recurrent tumors. No infiltrative feature was found in other 83 patients. Microscopic infiltrative growth patterns played an important role in predicting prognosis for the very-low-risk GISTs. No significant financial relationships to disclose.
Effect of the I1307K polymorphism in APC confers a higher risk for polyp recurrence in Jewish Ashkenazi carriers
Authors: Figer A. , Shacham-Shmueli E. , Liberman E. , Sagiv E. , Hall M. J. , Dolkart O. , Kazanov D. , Kraus S. , Neugut A. I. , Inbar M. , Arber N. 
Published: August 2020

e22003 Background: The I1307K adenomatous polyposis coli gene variant, prevalent among Ashkenazi Jews, may increase risk for colorectal neoplasia [colorectal cancer (CRC) and CR adenoma]. We studied the clinical importance of this polymorphism in 3283 Israelis undergoing colonoscopic assessment. Methods: Blood samples and risk factor information were collected from individuals undergoing colonoscopic examination at our medical center. Germline genetic analysis for the APC I1307K variant was performed using real-time PCR for DNA extracted from peripheral mononuclear cells. Results: The overall prevalence of the I1307K polymorphism was 8.1% (10.2% among Ashkenazi while only 2.5% among Sephardic Jews, p=0.001). The overall adjusted odds ratio (OR) for CR neoplasia among carriers was 1.3 (1.0 -1.7, p=0.049). Among Ashkenazi Jews, the I1307K variant was significantly more prevalent among persons with a personal or family history (1st degree) of CR neoplasia (p=0.01) as compared to Ashkenazi Jews with no family history. The histopathological features of adenomas and cancers did not differ between carriers and non-carriers. No interactions were found between the I1307K variant and demographic, lifestyle, or dietary modifiers that independently modulated the risk for CR neoplasia. Conclusions: In the general population, the APC I1307K variant does not change the risk or prognosis of colorectal neoplasia in carriers and does not necessarily change their clinical practice. Nevertheless, the variant, which is more prevalent among high risk individuals of Ashkenazi Jewish origin, is an important risk factor for the assessment of recurrence of neoplasia as it confers a higher risk for polyp recurrence in this population. No significant financial relationships to disclose.
The critical role of surgical pathology in personalized medicine: The impact of biopsy cavities in breast cancer samples on recurrence risk when assessed by quantitative RT-PCR
Authors: Baehner F. L. , Anderson J. , Millward C. , Sangli C. , Quale C. , Cherbavaz D. , Goddard A. , Shak S. 
Published: August 2020

e22016 Background: Tumor gene expression analysis using the Recurrence Score (RS) assay is frequently used in ER+ breast cancer. Manual microdissection is performed in cases where biopsy cavities (BxC) are present in the submitted specimen. The objective of this was to characterize by quantitative RT-PCR the impact of BxC on 21 gene expression profiles and the RS. Methods: 48 (15 well, 18 moderate, and 15 poorly differentiated) breast cancers were evaluated for gene expression differences between whole sections (WS; containing BxC) and enriched tumor (ET; BxC excluded). Standardized quantitative RT-PCR analysis for the 21 Oncotype DX genes was performed; reference normalized gene expression measurements ranged from 0 to 15, where each 1-unit reflects an approximate 2-fold change in RNA. Analyses of individual genes and the RS were performed on the entire sample set and stratified by tumor grade. Correlation analyses used Pearson's R, concordance analysis used Lin's sample concordance and paired t- tests to characterize differences. Results: There were statistically significant differences in reference normalized gene expression between ET and WS in 6 genes: BAG1 (ET-WS: 0.13 units, p=0.0025), CD68 (ET-WS: -0.64 units, p<0.0001), ER (ET-WS: 0.29 units, p=0.0012), GSTM1 (ET-WS: 0.18 units p=0.0025), STK15 (ET-WS: -0.18 units, p=0.0041) and STMY3 (ET-WS: 0.62 units, p<0.0001). Expression of the macrophage marker CD68 was higher and expression of ER was lower in WS containing BxC. The correlation (0.95) and concordance (0.92) were generally high between WS and ET for RS overall however among moderately differentially tumors, there was a statistically significant mean increase in RS for WS of 3.3 units (p = 0.0012) while among poorly differentiated tumors there was a trend toward a statistically significant decrease in RS for WS of 2.2 units (p=0.0569). Conclusions: Histologic identification of invasive carcinoma and exclusion of BxC is essential for precise RS assessment. Inclusion of BxC in breast cancer specimens is associated with significant changes in the expression of individual genes and impacts the RS. Removal of BxC from breast cancer specimens assessed for gene expression levels is warranted. [Table: see text]
Prognostic impact of polymorphisms of caspase genes on survival in patients with operated invasive ductal breast cancer
Authors: Park H. , Jung J. , Eun Y. , Kim J. , Chae Y. , Kim J. , Sohn S. , Park J. , Yang J. , Lee Y. 
Published: August 2020

e22020 Background: Since the apoptosis induced by caspases may play a role in the prognosis of breast cancer, the present study analyzed the polymorphisms of caspase genes and their impact on the survival in patients with operated invasive ductal breast cancer. Methods: Two hundred eighty patients with breast cancer who underwent surgery with curative intent were enrolled in the present study. The genomic DNA was extracted from paraffin-embedded tissue, and 7 single nucleotide polymorphisms (SNPs) of 6 caspase genes were determined using a PCR-RFLP assay. Results: The median age was 49 (range, 24–82) years, and 157 (56.1%) patients were premenopause at the time of diagnosis. Thirty-six (12.9%) patients underwent breast-conserving surgery. Pathologic stages after surgery were as follows: stage I (n=86, 30.7%), stage II (n=129, 46.1%), and stage III (n=63, 22.5%). Among the target SNPs, CASP6 (rs2301717, +5095G>T) and CASP7 (rs11593766, +11G>T) were found to correlate with relapse-free survival (RFS) after curative surgery in an univariate analysis adjusted to age, stage, histological grade, and the immunohistochemical expression of estrogen receptor and HER2. Meanwhile, multivariate analysis showed that the combined GT and TT genotype of CASP6 (rs2301717) was associated with a better RFS than the GG genotype (HR=0.39; 95% CI=0.19–0.79; p=0.009). However, there was no significant association with overall survival. Conclusions: The CASP6 (rs2301717) polymorphism may be considered as a possible prognostic factor for survival in patients with operated invasive breast cancer. No significant financial relationships to disclose.
Association of serum levels of the growth factor GP88 with disease progression in breast cancer patients
Authors: Serrero G. , Tkaczuk K. , Zhan M. , Tait N. , Ilan C. , Eklund D. , Yue B. 
Published: August 2020

e22021 Background: The autocrine growth factor GP88 is an important player in breast cancer. GP88 is expressed in human BC tumors in correlation with their tumorigenicity. Increased GP88 expression was associated with anti-estrogen therapy resistance in ER+ cells and Herceptin resistance in Her-2 overexpressing breast tumors. Inhibition of GP88 expression inhibited tumor incidence and growth in nude mice. Immunohistochemical studies have shown that GP88 is expressed in invasive ductal carcinomas (IDC) and that high GP88 expression correlated with increased recurrence and mortality. Since GP88 is found in serum, we hypothesized that GP88 was elevated in the sera of breast cancer patients compared to healthy individuals and that GP88 serum level increases with disease progression. Methods: An IRB approved prospective study was established at the University of Maryland Breast Clinic to determine the serum level of GP88 in breast cancer patients (BC pts). Approximately 5 ml of blood was drawn every three months. GP88 serum concentration was determined in triplicate by human GP88 enzyme immunoassay. 190 BC pts were accrued. Sera from healthy volunteers (HV) were obtained to establish GP88 baseline. BC patient characteristics: Caucasian- 91, African American-92, Asian-6; median age, 51 (range 29- 86), stage I - 48, II - 52, III - 26, IV - 63. Results: Median serum GP88 level was 28.7 ng/ml (range 16.6–38.2) in HV, 40.7 ng/ml (range 6.4–100) in early stage (stage 1 -3) BC pts (p- value = 0.007) and 45.3 ng/ml (range 9.8 to 158.4) in stage 4 BC patients (p- value= 0.0007). Statistically significant increase in serum GP88 level was found in early stages as well as in metastatic disease when compared to HV. In addition, patients that were initially diagnosed with early stage disease but recurred showed a 5 to 10 fold increase in their GP88 serum levels. Conclusions: GP88 serum level is significantly higher in the sera of BC than HV subjects. Moreover, GP88 serum level increased in association with disease recurrence and progression. This study identifies GP88 as a measurable biomarker for disease progression not only at the tissue but also at the serum level. These results are also interesting since GP88 is also a therapeutic target of malignant progression of breast carcinoma. No significant financial relationships to disclose.
Breast cancer patients stratification by microarray-based gene expression profiling from FFPET samples
Authors: Lih C. , Li Y. , Trinh L. , Chien S. , Wu X. , Liu W. , Williams P. M. 
Published: August 2020

e22041 Background: Microarrays have been used to monitor global genes expression and have aided the identification of novel biomarkers for patients stratification and drug response prediction . To date there has been limited application of microarray- based gene expression analysis to formalin fixed paraffin embedded tissues (FFPET). FFPE tissues are the most commonly available clinical samples with documented clinical information for retrospective clinical analysis. However, FFPET RNA has proven to be an obstacle for microarray analysis because of low yield and compromised RNA integrity. Methods: Using a novel RNA amplification method, Single Primer Isothermal Amplification (SPIA, NuGEN Inc, San Carlos, CA), we amplified FFPET RNA, hybridized amplified, and labeled cDNA onto Affymetrix HG U133plus2 GeneChips. Results: We found that SPIA amplification successfully overcomes the problems of poor quality of FFPET RNA, and produced informative biological data. Comparing the gene expression data from 5 different types of FFPET archival cancer samples (breast, lung, ovarian, colon, and melanoma), we demonstrated that gene expression signatures clearly distinguish the tissue of origin. Further, from an analysis of 91 FFPET samples comprised of ER+, HER2+, triple negative breast cancer patients, and normal breast tissue, we have identified a 103 gene signature that distinguishes the intrinsic sub-types of breast cancer. Finally, the accuracy of gene expression measured by microarray was verified by real time PCR quantitation of the ERBB2 gene, resulting in a significant correlation (R = 0.88). Conclusions: We have demonstrated the feasibility of global gene expression profiling using RNA extracted from FFPET and have shown that a gene expression signature can stratify patient samples into different subtypes of disease. This study paves the way to identify novel molecular biomarkers for disease stratification and therapy response from archival FFPET samples, leading to the goals of personalized medicine. No significant financial relationships to disclose.
Wnt signaling somatic alterations in apc-associated fap adenomas
Authors: Obrador-Hevia A. , Chin F. , Gonzalez S. , Vilardell F. , Cordero D. , Greenson J. , Moreno V. , Caldas C. , Capella G. 
Published: August 2020

e22046 Background: APC mutations are believed to constitutively activate the wnt pathway in colorectal tumors. Our goal was to comprehensively characterize Wnt signaling components in a set of APC-associated FAP tumors both at the DNA and RNA levels. Methods: Sixty adenomas from FAP cases harboring pathogenic APC mutations were included (10 adenomas and 1 normal mucosa per case). Somatic APC and KRAS alterations, β-catenin immunostaining and qRT-PCR of APC, MYC, AXIN2 and SFRP1 were analyzed. aCGH was also assessed in 26 FAP adenomas and 24 additional paired normal-adenoma-carcinoma samples. Results: A second APC alteration was present in 30 (50%) of adenomas (26 LOH and 4 point mutations). LOH was only occasionally associated with loss of genetic material. In 3 of 6 cases APC mRNA was overexpressed in macroscopically normal mucosa. In these cases diminished APC levels mRNA were observed in 11 of 30 adenomas analyzed.. In the remaining 3 cases APC mRNA was already underexpressed in normal mucosa with only 3 of 30 adenomas showing further underexpression. In FAP normal mucosae MYC was overexpressed (logratio range: 3.37–5.66). All adenomas showed further MYC (logratio range: 1.78–2.92) and AXIN2 overexpression (logratio range: 1.62–4.65), corregulating with APC mRNA levels (r=0.31 for MYC; r=0.59 for AXIN2). β-catenin nuclear immunostaining was detected in 80% of adenomas correlating with MYC mRNA levels. SFRP1 was underexpressed in all tumors (logratio range= -3.06–27). While copy number changes were rare in adenomas (median number :2.5; range 0–5). DNA changes were more often detected in areas containing wnt genes when FAP and sporadic tumors were jointly analyzed (p=0.02). Conclusions: Wnt pathway is altered, at the DNA and/or RNA level, in all APC mutant FAP tumors. MYC overexpression is a universal event that correlates with APC and AXIN2 expression levels as well as bcatenin nuclear accumulation. No significant financial relationships to disclose.
Genetic evolution of EGFR and the clonal origin of adenocarcinomas exhibiting various degrees of bronchioloalveolar carcinoma
Authors: Zhong W. , Yang X. , Guo A. , Su J. , Zhang X. , Chen H. , Qiao G. , Liao R. , Yang J. , Wu Y. 
Published: August 2020

e22050 Background: EGFR mutations may accumulate during multistage progression of bronchioloalveolar carcinoma (BAC), leading to heterogeneity within the tumor. Moreover, intrapulmonary emersions are the predominant sites of BAC progression in the absence of other distant metastases. In cases of emerging bilateral lung lesions during the follow-up to complete resection, the issue of how to differentiate between lesions originating from multifocal BACs or distant metastases/local recurrence is an important and unresolved issue. This study was performed to determine whether sequential adenocarcinoma with BAC features emerges in the lung field arises from a single clone or multiple clones in the same individual. Methods: Samples of adenocarcinomas exhibiting various degrees of BAC were obtained by thoracotomy. Sequential specimens were obtained on detection of novel lesions in the lung field. Genomic DNA was extracted from the specimens, and the presence of activating mutations in EGFR was analyzed by direct sequencing. Our pathological findings, sequential imaging, and EGFR sequence data were compared to monitor evidence of cancer evolution. Results: Based on an analysis of EGFR in tumor specimens from 428 lung cancer patients, fifteen cases of sequential BAC-related adenocarcinoma obtained by thoracotomy were identified. Together with alterations in BAC/adenocarcinoma components, the EGFR-TKI untreated series with at least one episode of EGFR-activating mutations represented three typical models: no significant EGFR evolution for a single clone, genetic alterations from mutant to wild-type EGFR for multifocal lesions, and a switch from wild-type to mutant EGFR, which might exhibit uncertain circumstances of cancer progression. Conclusions: Genetic analysis in conjunction with pathological and radiological diagnoses can be used to explore the origin of multifocal BAC. The single clone model indicates subsequent disease progression, whereas genetic alterations from mutations to wild-type EGFR are suggestive of secondary primary carcinoma. When additional lesions emerge after radical resection of BAC-related lung cancer, sequential tumor samples should be obtained for further evaluation. No significant financial relationships to disclose.
CD10 expression and risk of relapse in DCIS
Authors: Toussaint J. , Durbecq V. , Rouas G. , Altintas S. , Desmedt C. , Larsimont D. , Piccart M. , Sotiriou C. 
Published: August 2020

e22053 Background: DCIS is a heterogeneous malignant condition of the breast with no standard of care. The Van Nuys Prognostic Index (VNPI) is the most useful tool to predict local recurrence. However, assessment of the VNPI score is not easy and many women with DCIS are offered the same treatment. Experimental evidence has indicated that a local death of the myo-epithelial cells (MECs) in the DCIS is a pre-requisite to tumor invasion. In this study we hypothesized that loss of CD10, (a marker of MECs), would be a surrogate for basement membrane disruption and tumor invasion. The aim of the present study was to retrospectively evaluate the prognostic value of CD10 in DCIS. Methods: CD10 expression was evaluated by qRT-PCR and immuno-histochemistry (IHC) using FFPE tissues on normal samples (N=11) and two independent DCIS populations: a training set (N=66) and a validating set (N=88). Results: MECs were the only cells that showed a CD10 staining using IHC in normal and DCIS samples. All normal tissue samples demonstrated high expression levels of CD10 using both IHC and RT-PCR. In contrast, DCIS samples showed a wide range of CD10 expression levels. Of interest, 66% of DCIS showed lower expression values of CD10 compared to normal tissues. The group of DCIS with low expression of CD10 was statistically associated with higher risk of relapse [HR: 2.49 (CI.95%= 1.13–5.49), P= 0.02]. These results were independently validated in our validation set using both techniques (qRT-PCR: HR: 1.805 (CI.95%= 1.11–2.92), P= 0.02]; IHC [HR: 1.84 (CI.95%= 1.00–3.40), P= 0.05]. In multivariate analysis, CD10 remained significant (HR=2.25; [95CI: 1.24–4.09], p=0.008) together with VNPI [HR: 2.03 (CI.95%= 1.23–3.35), P= 0.006]. Conclusions: Decrease of CD10 expression in MECs is associated with higher risk of relapse in DCIS and it has the potential to improve DCIS management. These promising results are currently being validated in a larger patient's series. No significant financial relationships to disclose.