bluuebunny/crossref_metadata_2025 huggingface dataset:
Use of Port-A-Cath in pediatric cancer patients: Experience from a tertiary cancer center in south India
Authors: Singh T. , Satheesh C. , Ankit J. , Sajeevan K. V. , Appaji L. , Arunakumari B. , Padma M. , Mamatha H. S.
Published: August 2020

e20747

Background: Implanted subcutaneous (s.c.) central venous port accesses including Port-A-Cath (PAC) facilitate the administration of chemotherapy or blood products and are frequently used in children with cancer. We present our experience of PACs in paediatric cancer patients from a tertiary cancer centre in South India. Methods: A total of 20 patients with paediatric malignancies requiring PAC were included in the study. Aim was to review our experience of PAC and analyse the outcome in paediatric cancer patients in Indian setting. A record of all complications and final outcome were analysed. Results: A total of 20 PAC were placed. Disease distribution included ALL(80%), AML(5%), NHL(5%), neuroblastoma (5%) and RMS (5%). PAC has been in place for 2 to 36 months (cumulative 400 months). Portal infection (Coagulase-negative staphylococci) was observed in four patients of whom 3 patients had their PAC removed (portal occlusion also seen in one of the pt). Conclusions: Although, catheter-related infections demanded PAC removal in 15% of our patients, their benefits (excluding cost 300 $) argue for continued PAC use in the paediatric cancer population as it is safe and has many advantages compared to traditional CVCs in use. Strict indications, meticulous implantation technique, and adequate handling are, however, mandatory.

No significant financial relationships to disclose.
2008 oncology trends: Views from payors, specialty pharmacy, and oncology providers
Authors: Holcombe D. G.
Published: August 2020

e20750

Background: Kikaku International Research was engaged by Genentech USA to conduct survey research among four key stakeholder groups involved in the financing, management and delivery of cancer care and drugs. Methods: Surveys were distributed and collected from January to June 2008. Survey questions addressed issues on volume of services, utilization of specialty pharmacy, management methodologies and degrees of involvement, volume and staffing levels in oncology practice, oncology physician staffing and retirement, payor evaluations, cost and impact of public and private payor reimbursement policies, Average Selling Price (ASP) and Medicare Modernization Act (MMA) issues and use of guidelines/pathways. Results: Managed Care Organizations: Ninety (90) managed care professionals responded. Key findings addressed: anticipated shifts in cancer spending from the medical benefit to the pharmaceutical benefit, adoption of ASP-based reimbursement and revaluation of professional fees. Specialty Pharmacy Providers: Forty-two (42) specialty pharmacy professionals responded. Key findings: total revenues and volume from cancer drugs and supportive care, distribution of drugs to patients and physician offices, Injectable drugs and oral drugs precription volume, patient education and disease management. Oncologists - Surveys were returned by 139 oncologists. Key findings: workloads and patient loads, income declines, drug margins declines, buy and bill volume. Oncology Practice Managers: Sixty (60) oncology practice managers responded to surveys about practice management, billing and reimbursement. Key findings: payer coverage and reimbursement policies, ASP rates, health plan contracts, negotiation of fee schedules. Conclusions: The conclusions of each section of the survey, when combined in aggregate, paint a picture of a very complex topic with wide variation in both approach and recommendations for how each segment addresses the specialty. Disconnects, challenges and opportunities observed in evaluating the responses include perspectives on ASP, Medical vs pharmaceutical benefit, contract negotiations, disease management, pharmaceutical management.

[Table: see text]
Proliferation heterogeneity in synovial sarcoma (SS) defines different patterns of clinical outcome: A retrospective study of 32 patients
Authors: Grisanti S. , Consoli F. , Ferrari V. D. , Bercich L. , Amoroso V. , Rangoni G. , Cetto G. L. , Dei Tos A. P. , Facchetti F. , Marini G.
Published: August 2020

e21511

Background: SS is an aggressive soft tissue sarcoma (STS) characterized by a constitutive overexpression of the bcl-2 proteins and high proliferation rate (Ki67). The aim of the study was to evaluate prognostic value of proliferative activity and apoptosis in SS. Methods: A retrospective analysis of 32 patients treated at three oncology centers between January 2000 and August 2008 was conducted. Histologic diagnosis of SS was confirmed by FISH analysis of t(X;18). Bcl-2 and Ki67 were determined by immunohistochemistry at baseline and after neoadjuvant chemotherapy (CT). A cut-off value of 20% was established for Ki67. The bcl-2 gene status was evaluated by FISH in neoadjuvant subgroup. Treatment-induced pathological response (pCR) was defined as tumor necrosis of 100%. Endpoints were recurrence rate (RR), disease-free survival (DFS) and overall survival (OS). Clinical and pathological variables were considered in uni- and multivariate analysis. Results: 13/32 patients received an anthracycline/ifosfamide-based chemotherapy (CT) before surgery. 8 pts received concomitant radiotherapy. A median number of 3 cycles of neoadjuvant CT was administered. All patients underwent surgical resection and pathologic assessment of the coexpression of bcl-2 and Ki67 was evaluated in the resected specimens. At baseline, all samples showed immunoexpression of bcl-2 and 9/13 had Ki67 more than 20%. After neoadjuvant CT, Ki67 was downregulated <20% in 11/13 samples and bcl-2 was negative in 4/13 pts. At FISH analysis, bcl-2 gene was neither rearranged nor amplified. After a median follow-up of 21 months (range 13–64 months), 9/13 pts without pCR experienced progression disease and 5 of them were dead of SS. At uni- and multivariate analysis, both pCR and downregulation of bcl-2 and Ki67 activity had a significant impact on DFS and OS (p=0.03). Conclusions: 1. Downregulation of bcl-2 and Ki67 index after treatment could be a predictor of recurrence and overall survival. 2. SS is characterized by cellular heterogeneity in which a high proliferative compartment coexists with low proliferative/anti-apoptotic compartment with different response to treatment.

No significant financial relationships to disclose.
Management of complicated tumor response (CTR) to tyrosine kinase inhibitors (TKI) in gastrointestinal stromal tumor (GIST) patients (pts) with locally advanced/metastatic disease
Authors: Coco P. , Fumagalli E. , Morosi C. , Messina A. , Bertulli R. , Stacchiotti S. , Gronchi A. , Casali P.
Published: August 2020

e21512

Background: Bleeding, perforation, and infection may complicate TKI therapy in GIST. Their incidence and best management are not completely known yet. Methods: Between May 2001 and December 2008, 278 pts were treated with imatinib or sunitinib, respectively, as first or second line therapy. Pts experienced bleeding, perforation and colliquation with superimposed infection were recorded and reviewed. Results: CTR occurred in 24 pts (9%) after a median time of 241 days from treatment start (8–2090 days). In 5 pts the complication took place after increasing the dose. Before complication, according to RECIST criteria, a partial response (PR) and a stable response (SD) were respectively documented in 16 and 8 pts . Radiologically assessed bleeding in closed lesions was seen in 13 pts (5%). All but one were treated conservatively with supportive care and blood transfusions. The other pt required surgical resection of his peritoneal bleeding lesion. Nine pts (3%) developed colliquation with bacterical superinfection: 5 were treated with antibiotics, 4 had their lesions drained. Two pts underwent emergency surgery due to perforation with a superimposed infection. In 21 out of 24 pts the event resolved without sequelae. Three pts died: 2 developed massive haemorrhage, 1 had a bowel perforation. After stopping treatment for a median interval of 10 days (2–63), all but one restarted treatment at the previous dose. No further complications were seen after the first event. Eight pts are still on treatment with a SD, 2 pts switched to a second line therapy due to progressive disease (PD), 11 died due to PD. Conclusions: Bleeding, perforation and colliquation with superimposed infection may occur at different time intervals from treatment start. Even if supportive care and medical therapy are successful in most of cases, pts should be carefully monitored because these complications can be life-threatening. Surgery should be considered in case of failure of conservative treatment. Reintroduction of the therapy at the same dose is feasible and well tolerated.

[Table: see text]
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.
Nuclear factor-kB (NF-kB), p53, survivin, Ki-67, and Bcl-2 as prognostic factors in locally advanced rectal cancer patients receiving radiochemotherapy
Authors: Berardi R. , Mandolesi A. , Onofri A. , Maccaroni E. , Mantello G. , Bearzi I. , Scartozzi M. , Pierantoni C. , Marmorale C. , Cascinu S.
Published: August 2020

e22062

Background: NF-kB, p53, Survivin, Ki-67 and Bcl-2 expressions have been demonstrated to be prognostic factors in solid tumors. The aim of our analysis was to investigate the importance of their expression, as prognostic factors in patients with locally advanced rectal cancer patients receiving receiving neoadjuvant radiochemotherapy Methods: We analyzed the expression of NF-kB, p53, Survivin, Ki-67 and Bcl-2 in patients with locally advanced rectal cancer who underwent neoadjuvant treatment (radiotherapy ± chemotherapy) at our Department Results: Seventy-four patients were eligible for our analysis. Median age at diagnosis was 66 years (range 36–85). Male/female ratio was 47/37; 37 patients (90%) were diagnosed with adenocarcinoma, whilst 4/41 (10%) with mucinous adenocarcinoma. All the patients received radiotherapy ± 5-fuorouracil/capecitabile-based chemotherapy. Median follow up was 28 months (range 6,7–56,6 months). At univariate and multivariate analysis of the above mentioned parameters, NF-kB, Ki67 and bcl-2 showed an impact on outcome.In particular, in NF-kB-strongly positive patients time to progression (TTP) and overall survival were significantly shorter (p=0,011 and p=0,018 respectively). Moreover a high expression of Ki-67 and a low expression of bcl-2 were associated with a better TTP Conclusions: Our results suggest that NF-kB, bcl-2 and Ki-67 could represent important parameters able to predict the outcome in patients receiving neoadjuvant treatment for rectal cancer. Further prospective studies are warranted in order to confirm the prognostic role of the above mentioned factors in this setting. This could be useful in order to select patients to receive adjuvant chemotherapy after neoadjuvant treatment and surgery for locally advanced rectal cancer, intensifying the adjuvant therapy in some groups of patients and obviating the use of the some drugs (i.e. those involving NF-kB in their mechanism of action) in selected patients

No significant financial relationships to disclose.
Differential outcomes in patients treated with endocrine therapy for early or locally advanced breast cancer based on BRCA mutation status
Authors: Wesolowski R. , Shealy A. G. , Tao J. , Moore H. C.
Published: August 2020

e22065

Background: Mutations in BRCA1 and BRCA2 genes lead to defects in DNA repair. Estrogen receptor modulates transcription of genes responsible for cell division, which depends on cell's ability to repair DNA for genomic integrity. Differential efficacy of endocrine therapy for breast cancer, therefore, may be possible depending on the tumor's BRCA mutation status. Methods: Through an IRB approved registry, breast cancer patients tested for BRCA1 and BRCA2 mutations and treated with endocrine therapy for hormone-receptor positive non-metastatic disease were identified. Primary and secondary endpoints were overall survival (OS) and progression-free survival (PFS) respectively. Fisher's exact test or Wilcoxon rank sum test were used to assess differences among subgroups with respect to their characteristics. Cox proportional hazard analysis was used to identify univariate and multivariate risk factors for OS and PFS. Results: Of 115 breast cancer patients tested for BRCA mutations, 63 met the inclusion criteria of whom 16 patients were BRCA 1 or 2 mutation positive and 47 were negative. In the BRCA(+) group, 14 patients (87.5%) had stage I-III disease at diagnosis. In the BRCA(-) group, 5 patients (10.6%) had stage 0 disease while 41 patients (87.2%) had stage I-III disease at diagnosis. Stage at diagnosis was unavailable for 2 BRCA(+) and 1 BRCA(-) patients. Both groups were similar with respect to Her-2 expression status, history of ovarian suppression, age of diagnosis, and age of menopause. Median age was 48 yo in BRCA(+) group, 42 yo in BRCA(-), (p=0.12). Median follow up was 76.1 mos in BRCA(+) and 62.9 mos in BRCA(-) group. OS was worse in BRCA(+) group (HR 7.38, 95% [CI] 1.49–36.4 p=0.014). After adjustment for stage and history of ovarian suppression, the difference remained significant (HR 16.6, 95% [CI] 1.95–142, p=0.010). There was no difference in PFS (HR 2.02, 95% [CI] 0.82–4.96, p=0.13). Conclusions: Patients with BRCA mutation, hormone-receptor positive hereditary breast cancer treated with endocrine therapy had inferior survival compared with similar patients who are BRCA mutation negative. Prospective studies to evaluate the differential effects of endocrine therapy in these populations are warranted.

No significant financial relationships to disclose.
Molecular changes in epidermal growth factor receptor (EGFR) in non-small cell lung cancer (NSCLC) biopsies at time of progression compared to initial biopsy
Authors: Speranza G. , Cohen V. , Agulnik J. S. , Chong G. , Meilleur F. , Brandao G. , Kasymjanova G. , Small D. , Miller W. H.
Published: August 2020

e22066

Background: EGFR mutations predict sensitivity and clinical outcome to tyrosine kinase inhibitors (TKI) in NSCLC. The two most commonly described mutations are Exon 19 deletion and Exon 21 L858R missense mutations. Genetic alterations over time have been described in other tumour types, but studies assessing EGFR genotypic changes with lung cancer progression are lacking. We sought to compare EGFR mutational status from lung tumors at time of recurrence or progression with the primary tumor. Methods: Using the Jewish General Hospital lung cancer database, of all patients diagnosed with NSCLC since 1999, those with biopsies at two different points in time were identified. All tumour samples were genotyped for EGFR exons 19 and 21 mutations using denaturing high performance liquid chromatography (dHPLC). Results: 29 patients were identified. Data for 12 patients, whose time of recurrence or progression varied between 4 months and 6 years, are available at this time. Of 12 patients, one had EGFR exon 19 mutation at time of diagnosis. One patient who initially displayed no EGFR mutation was found to have an exon 19 deletion at time of recurrence. The one with exon 19 at time of initial diagnosis continued to express exon 19 in the second biopsy. Conclusions: To our knowledge, this is the only study assessing changes in molecular genotype using dHPLC between primary and recurrent or progressive lung cancer biopsy specimens. Although sample size is small, it is evident that changes in EGFR mutational status can occur. Further prospective studies are required to determine how commonly molecular changes occur.

No significant financial relationships to disclose.
MicroRNA-based assay for differential diagnosis of mesothelioma
Authors: Benjamin H. , Lebanony D. , Tabak S. , Barabash N. , Gibori H. , Morgenstern S. , Meiri E. , Bentwich Z. , Rosenwald S. , Cohen D.
Published: August 2020

e22079

Background: Malignant mesothelioma is an aggressive pleural neoplasm, strongly linked to environmental exposures such as asbestos. Mesothelioma can be difficult to differentiate from other tumors in the lung or pleura such as primary lung adenocarcinoma presenting with pleural effusion or metastatic adenocarcinoma from extrathoracic sites. We addressed the increasing need for accurate differential diagnosis of these tumors by developing a diagnostic assay based on expression levels of microRNAs, a family of small, non-coding RNAs whose tissue-specificity has proven applicability for identification of cancer tissue type and histology. Methods: We developed protocols for extraction of high-quality RNA that retain the microRNA fraction from FFPE tissue samples. Microarrays were used for initial profiling. qRT-PCR was used to validate results and to develop a diagnostic assay. Results: We identified microRNAs that are differentially expressed between mesothelioma, lung adenocarcinoma, and other confounding tumor types. A diagnostic assay (miRview™ meso) was developed, that utilizes qRT-PCR measurement of a small set of microRNAs to differentiate between mesothelioma and non-mesothelioma samples. After establishing this profile in more than 30 mesotheliomas and 200 samples of confounding tumors, the microRNA biomarkers were measured using a standardized protocol on a blinded test set. The assay had accuracy greater than 90% in differentiating mesothelioma from other confounding tumor types. More than ¾ of samples were classified with high confidence, and these samples were all correctly identified. Conclusions: MicroRNAs are emerging as effective cancer biomarkers. A robust and simple assay based on the expression level of a few microRNA biomarkers can accurately differentiate mesothelioma from other possible tumors in the lung and pleura. This assay provides an important new tool for diagnosing mesothelioma.

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Use of multidrug-resistance gene (MDR1) expression levels to predict outcome to adjuvant cisplatin/gemcitabine based chemotherapy in patients with with locally advanced urothelial cancer
Authors: Gauler T. , Hoffman A. C. , Wild P. , Leicht C. , Bertz S. , Danenberg K. D. , Stöhr R. , Stöckle M. , Eberhardt W. E. , Lehmann J. , Hartmann A.
Published: August 2020

e22081

Background: The human MDR1 gene encodes an integral membrane protein, P glycoprotein (Pgp), whose function is the energy dependent export of substances from the inside of cells, and from membranes, to the outside. MDR1 mRNA levels have been shown to influence metabolism of cancer drugs. We tested whether MDR1 gene expression is associated with outcome in patients with locally advanced bladder cancer. Methods: 43 formalin fixed paraffin embedded (FFPE) tumor samples from patients with locally advanced and/or lymph node-positive bladder cancer undergoing treatment with cisplatin/gemcitabine were analyzed.FFPE tissues were dissected using laser-captured microdissection and analyzed for MDR1 expression using a quantitative real- time RT-PCR method. Gene expression values (relative mRNA levels) are expressed as ratios between the target gene and internal reference gene (beta-actin). Results: MDR1 was significantly correlated to overall survival. We included tumor stage (pT), lymph node status (pN) and the blood vessel invasion along with the measured gene expression in a stepwise multivariate Cox proportional hazards regression model. The overall model fit had a significance level of p=0.04 (<0.05). The relative risk for shorter survival in patients with High MDR1 expression was 1.9. Conclusions: These results suggest that MDR1 gene expression levels predict response and overall survival in patients with locally advanced urothelial cancer. MDR1 gene expression levels may allow the selection of patients who benefit likely from adjuvant cisplatin/gemcitabine based chemotherapy. Further studies are warranted to study this association.

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The role of HER3 on HER2-driven gallbladder carcinogenesis and possible HER2/HER3 targeting therapy by pertuzumab for gallbladder canrcinoma
Authors: Kawamoto T. , Ishige K. , Sugiyama H. , Onuki K. , Krishnamurthy S. , Thomas M. , Yamamoto M. , Shoda J.
Published: August 2020

e22083

Background: ErbB2 has been known to be important for gallbladder carcinogenesis. Supporting this, the gene amplification of HER2 was found in ca. 20% of human gallbladder carcinoma (GBC) (Kawamoto, et al. Gastrointest Cancer Res 2007). Following these results, a Phase II study of trastuzumab for treatment of GBC (NSC 688097) is being conducted. Although the mechanism of HER2 activation is not well understood, the role of HER3 in HER2-driven tumorigenesis has been discussed and we have concluded that HER2/HER3 heterodimerization might be important for GBC cell proliferation (ASCOGI 2009, ab#155). In this study, we determined HER family expressions in human GBC and examined the effect of pertuzumab against human GBC cell lines. Methods: Tissues from 47 GBCs and 6 non-cancerous gallbladders were examined. All cases were screened for HER1, HER2, HER3 expressions by immunohistochemistry (IHC) and those HER family gene amplification by fluorescence in situ hybridization (FISH) were performed. 6 human GBC cell lines were also analyzed by Western blotting and FISH as well and their growth assays were investigated with heregulin stimulation to confirm the existence of HER2/HER3 heterodimerization. Then, GBC cells were cultured with heregulin and various doses of pertuzumab for 72 hours and CCK-8 assay was performed. Results: HER3, HER2 and HER1 overexpression by IHC was found in 34%, 32% and 19% of GBCs, respectively. Phosphorylated (p-) HER2 and p-HER1 were found in 23% and 11% of GBCs, respectively. FISH analysis was considered successful in the same serial sections. HER3, HER2 and HER1 FISH (+) was found in 26%, 19% and 4% of GBCs, respectively. Three of 6 GBC cell lines were recognized their growth curves with heregulin were increasing. These 3 cell lines showed good responses to the treatment with pertuzumab. Conclusions: More than 20% of GBC over-expressed either HER2 or HER3. These HER2 and HER3 may form heterodimer, which in turn results in the activation of HER2. The cell lines proliferating with the administration of heregulin showed good responses to the pertuzumab treatment. Therefore, HER2/HER3 heterodimerization may be an ideal biomarker for the treatment by pertuzumab and this agent may be a new therapeutic regimen for GBC.

No significant financial relationships to disclose.
Immunohistochemical analysis of tumor regression grade for rectal cancer after neoadjuvant chemoradiotherapy
Authors: Moreno Garcia V. , Cejas P. , Feliu J. , De Castro J. , Belda-Iniesta C. , Barriuso J. , Larrauri J. , Gonzalez Baron M. , Casado E.
Published: August 2020

e22089

Background: Tumor regression grade (TRG) as defined by Rodel et al. has been suggested as an independent prognostic factor for rectal carcinoma patients treated by preoperative chemoradiotherapy (CRT). However TRG 2 and 3 determination, semiquantitatively defined as more/less than 50% tumor regression, respectively, do not clearly correlate with prognosis. TRG 2 vs 3 discrimination is largely subjective hurdling prognostic analysis. The purpose of our study was to find an immunohistochemical pattern to better stratify these patients according to prognosis in term of disease free survival (DFS) and overall survival (OS). Methods: Immunohistochemistry of EGFR, VEGF, CD133, p53 and Ki67 was evaluated by tissue microarrays on surgical specimens from 88 patients. Preoperative chemotherapy was UFT-LV (30%) or oxaliplatin-based (70%) plus pelvic radiotherapy (50 Gy) followed by mesorectal excision. TRG was determined by the amount of viable tumor versus fibrosis, ranging from TRG 4 when no viable tumor cells were detected, to TRG 1 when regression was less than 25%. TRG 2 and 3 were defined as described above. Univariate analyses were performed according to the Kaplan-Meier method. Comparisons between curves were evaluated by the log-rank test. Cox regression was used for the multivariate analysis. Results: At a median follow-up of 39 months, the TRG was an independent predictor of DFS (p=0.05) and OS (p=0.001) but no differences were found between TRG 2 and 3 in terms of DFS (p=0.74) or OS (p=0.41). Our results show an immunohistochemical bad prognostic profile for tumors TRG 2 and 3 configured by double negativity of EGFR and CD133 expression (less than 5% of tumor cells membrane immunoreactivity for both antibodies). 3-year DFS and OS for these patients (vs. TRG 2 and 3 not-double negative) were 33 vs 65% (p=0.05) [HR=2.4 (95%CI, 0.9–6.1) p=0.06] and 77 vs 95% (p=0.06) [HR=4.1 (95%CI, 0.7–21.5) p=0.09]. Conclusions: The EGFR/CD133 double negative rectal tumors with TRG 2 or 3 after chemoradiotherapy show a higher risk of relapse or death. These results can help clinicians to determine better individual prognosis and are worth to confirm prospectively.

No significant financial relationships to disclose.