bluuebunny/crossref_metadata_2025 huggingface dataset:
High-throughput analysis of DNA repair in microplates towards identification of inhibitors
Authors: Kamiya Hiroyuki
Published: March 2020

Environmental factors can inhibit DNA repair and cause indirect mutagenic actions. Here the author introduces a recent paper on a novel high-throughput system to analyze the various enzymatic activities involved in base excision repair. Such systems will facilitate the identification of compounds that suppressively affect DNA repair.
Frameworks for health systems performance assessment: how comprehensive is Ghana’s holistic assessment tool?
Authors: Kumah Emmanuel , Ankomah Samuel E. , Fusheini Adam , Sarpong Emmanuel Kusi , Anyimadu Eric , Quist Ato , Koomson Brian
Published: March 2020

Background

Measuring the performance of a health system is an essential requirement in creating systems that generate efficient, equitable, patient-focused, accessible and sustainable results. A fundamental requirement for a performance measurement system is the development of an assessment framework within which specific performance measures could be defined and applied regularly. This paper examines the comprehensiveness of Ghana’s health system assessment framework called the Holistic Assessment Tool in relation to some of the internationally recognized frameworks. The paper also analyzes trends in the performance of the health system to understand whether or not an improvement has been recorded following the adoption and implementation of the Holistic Assessment Tool.

Methods

Mainly secondary data were used in this analysis. Searches were conducted on Google Scholar, PubMed, Scopus and Science Direct between May and July, 2019 for published documents on health system performance assessment. We also obtained unpublished documents from Ghana’s Ministry of Health, Ghana Health Service website, and Ghana Statistical Service database. Descriptive statistics were used to examine trends in the performance of the Ghanaian health system.

Results

While the tool provides a national framework for evaluating the performance of the Ghana Health system in several domains, the Holistic Assessment Tool does not cover key health system domains such as information systems for health, access to essential medicines, and patient-centeredness. Also, the scope of the assessment program seems limited to the evaluation of the Ministry of Health’s annual plans, programs and projects. However, the health system has recorded improvements in population health indicators, such as life expectancy at birth, infant mortality, under-5 mortality, HIV prevalence and disease burden (in terms of disability adjusted life years).

Conclusions

The Holistic Assessment Tool is a useful framework, but needs further refinement, both in scope and in conceptual robustness. Future studies should consider exploring factors influencing performance of the Ghanaian health system. Such information will help in strategizing for better and more improvements.
Nucleotide excision repair polymorphisms modulate overall lung cancer survival and responsiveness to platinum based chemotherapy agents
Authors: Matakidou A. , Eisen T. , Bridle H. , Houlston R. S.
Published: March 2020

10004

Background: The role of DNA repair in maintaining genomic stability and its role in modulating response to chemotherapy agents raises the possibility that variation in these genes may affect lung cancer prognosis. Methods: To comprehensively evaluate the impact of variation in the nucleotide excision repair (NER) pathway on lung cancer prognosis, we analysed 408 lung cancer patients for all coding, non-synonymous single nucleotide polymorphisms (SNPs) within NER-related genes. Results: Mean age of lung cancer diagnosis among patients was 63 years, 105 patients with small cell lung cancer (SCLC) and 299 with non-small cell lung cancer (NSCLC). The mean follow-up time was 24 months and a total of 234 deaths were observed. Mean survival time was 17.3 and 18.6 months for SCLC and NSCLC respectively. The XPG H1104D minor allele homozygote and the tightly linked CSB M1097V, R1213G, and Q1413R heterozygote genotypes were associated with decreased overall survival, irrespective of whether patients were treated with or without platinum chemotherapy (HRs of 2.22, 95% CI: 1.19–4.16; p=0.013 and 1.39, 95% CI: 1.05–1.84; p=0.02 respectively). No statistically significant interaction was detected between the at-risk genotypes and histology and/or stage of disease at diagnosis. To assess the predictive value of these SNPs in relation to platinum based chemotherapy, genotype-survival associations were evaluated in non-platinum and platinum treated patients. In non-platinum treated patients the RAD23B A249V and CSB M1097V minor allele genotypes were associated with decreased overall survival (p=0.004 and 0.002 respectively), whilst the XPC K939Q minor allele genotype was predictive of improved survival (p=0.03). In platinum-treated patients, carriers of the XPG H1104D minor allele were found to have a statistically significant decrease in their overall survival (p=0.04 and 0.004 for minor allele heterozygote and homozygote patients respectively). Conclusions: These findings suggest that genetic polymorphisms in the NER genes may modulate overall lung cancer prognosis as well as response to platinum-based chemotherapy agents, independent of histological diagnosis and stage of disease at presentation.

No significant financial relationships to disclose.
Analysis of genomic DNA copy number alterations in chromosome arm 18q demonstrates distinct molecular categories of colorectal carcinoma
Authors: Ji H. , Zhang M. , Farnam K. , Salari K. , Davis R. , Ford J. M.
Published: March 2020

10005

Background: Genomic instability is a major feature of neoplastic development in colorectal carcinoma and other cancers. Specific genomic instability events such as genomic deletions have potential utility as biologically relevant prognostic biomarkers. Loss of heterozygosity (LOH) on chromosome arm 18q is an indicator of colorectal carcinoma behavior and potentially, prognosis. For stage II and III colorectal cancer, a number of retrospective studies have shown strong correlations between deletions in 18q and reduced survival. However, other studies have failed to identify this correlation. This discrepancy is likely the result of different sets of genetic markers and their widely disparate locations along chromosome arm 18q, often separated by megabases. New technologies are needed to overcome these issues. Methods: Using a novel genomic technology called molecular inversion probes (MIPs), we analyzed genomic deletions at exon-level resolution in primary colorectal carcinoma. Unlike microsatellite genetic markers, MIPs are not dependent on having informative alleles to discern a LOH event. This enables querying microsatellite genetic markers used in any clinical study. We designed a set of probes to interrogate several hundred individual exons of over two hundred cancer genes with an overall distribution covering all chromosome arms. Also represented were a series of over 100 probes along chromosome arm 18q representing genetic markers used in clinical studies. An analysis was carried out on primary tumor samples from patients with stage II and III disease. Results: We discovered several distinct categories of colorectal carcinomas based upon their profile of 18q genomic deletions and other allelic imbalances. Colorectal carcinoma extent of invasion showed an association with cluster designation which suggests that the multiple genomic instability events influence the invasive behavior of colorectal carcinoma. Conclusions: Colorectal cancer has distinct patterns of 18q genomic deletions, representing a potential molecular classifier. This finding has potential clinical ramifications given the application of 18q LOH events as an indicator for adjuvant treatment in stage II colorectal carcinoma.

No significant financial relationships to disclose.
Phase II breast cancer chemoprevention trial of the third generation selective estrogen receptor modulator arzoxifene
Authors: Fabian C. J. , Kimler B. F. , Anderson J. R. , Chamberlain C. , Mayo M. S. , Zalles C. M. , O’Shaughnessy J. A. , Lynch H. T. , Johnson K. A. , Browne D.
Published: March 2020

1001

Background: Arzoxifene (ARZ) is a third generation SERM with efficacy in metastatic breast cancer but lacking uterine agonist activity. Methods: We conducted a randomized, double-blind, placebo-controlled Phase II prevention trial in 199 high risk women assessing the effects of ARZ 20 mg/day on several risk biomarkers. Biomarkers, including cytomorphology of breast epithelial cells obtained by random periareolar FNA (RPFNA) were assessed at baseline and following 6 months of placebo or ARZ. Subjects were stratified by presence or absence of atypia, ER expression, BRCA1/2 mutation, as well as menopause status. Results: At entry, mean age was 47, 52% were premenopausal and 47% of postmenopausal women were taking HRT. Mean 10 year Gail was 6.8% and mean Masood cytology index score was 14.3. The ARZ and placebo groups were well matched. The protocol defined primary endpoint was a decrease in RPFNA cytology Masood index score by ≥3 points at 6 months and required 160 evaluable subjects for 81% power to detect a change from 30% to 52% of subjects showing improvement. For the 181 evaluable subjects, there was no significant difference in the proportion of women achieving ≥3 point improvement (19% placebo vs. 24% ARZ, p=0.46); or in change in mean index score (0.6 placebo vs. 0.9 ARZ, p=0.53). There was also no difference in grade 3 or 4 side effects or dropout prior to 6 months. However, comparing ARZ to placebo, there was favorable modulation of the two risk biomarkers, mammographic breast density (p=0.001) and IGF-1:IGFBP-3 ratio (p=0.001), and reduction in bone turnover biomarker osteocalcin (p= 0.002), but without an increase in endometrial thickness. Conclusions: Although improvement in cytomorphology after 6 months of ARZ was not shown, the acceptable side effect profile and favorable modulation of other biomarkers (breast density, IGF-1:IGFBP-3, osteocalcin) provides support for continued evaluation of ARZ as a breast cancer prevention agent.

No significant financial relationships to disclose.
Case control study of hormone replacement therapy (HRT) and breast cancer in Japanese women
Authors: Takeuchi M. , Saeki T. , Sano M. , Sonoo H. , Honjo H. , Ochiai K. , Sugano H. , Komiike Y.
Published: March 2020

10012

Background: Some prospective studies in US and UK (represented by Women’s Health Initiative and Million Women Study) show that HRT use may increase the risk of breast cancer. However, it is unclear whether or not this perspective is applicable to Japanese women, due to several factors such as the difference in incidence of breast cancer, difference in HRT prevalence, which are to be concerned in this study. To investigate the relationship between HRT use and breast cancer, a case-control study was conducted in Japan. Methods: We sent self-administered questionnaire to 4,500 cases who have a previous history of confirmed histological breast cancer and 4,500 controls who were selected according to the inclusion criteria. All the subjects were between age 45 and 69 at the time of enrollment. Among these subjects, 3,434 in case group and 2,427 in control group sent back their questionnaire (total 5,861). The datacenter was run in Kitasato Univ. and the statistical analysis was performed using SAS (version 9.1). Control was selected by considering the screening record for GI disease and respiratory disorders, with no previous history of breast cancer, gynecological and hormonal disease at the time of screening. We asked about past history of exposure to the factors supposed to be breast cancer-causing; previous or current use of HRT, age at diagnosis, academic background, BMI, lifestyle habits, age of menarche, birth history, history of breast feeding, family history, use of contraceptive agents and menopausal status. Results: As the main analysis result, 164 (5.0%) out of 3,316 cases and 253 (7.4%) out of 2355 controls had used HRT (Odds ratio 0.432 [95% CI 0.352–0.530]. Conclusions: The result of this study shows that HRT users were less likely to develop breast cancer than never users. However, in this study, there was a difference in the observation period for the past use of HRT between cases and controls. Considering the transition of HRT prevalence in Japan, there may be a possibility that this difference can be a cause of a serious bias for the main analysis result. Further sensitivity analyses are needed to evaluate the robustness of the findings and this point is under consideration.

No significant financial relationships to disclose.
PTEN activity could be a predictive marker of trastuzumab efficacy in the treatment of ErbB2-overexpressing breast cancer
Authors: Fujita T. , Kawasaki K. , Takabatake D. , Takahashi H. , Ogasawara Y. , Doihara H.
Published: March 2020

10026

Background: Trastuzumab is the only HER2/neu-directed therapy for the treatment of patients with metastatic breast cancer. The efficacy of trastuzumab depends on the HER2/neu status of the tumor and the patient’s prior treatment, but even when patients are selected on the basis of HER2/neu gene amplification, the single-agent response rate ranges from 12 to 30% and few patients respond to trastuzumab monotherapy. Here we propose PTEN as a predictive biomarker for trastuzumab efficacy. Methods: Human breast cancer SKBR3 and drug-resistant SKBR3/R cells were analyzed, in vitro. Also we examined, retrospectively, clinical samples from patients who had been treated with trastuzumab for the metastatic disease and analyzed the relationship between trastuzumab efficacy and PTEN expression profile with immunihistochemistry. The PTEN expression level was scored semiquantitatively based on staining intensity and distribution using the immunoreactive score. Statistical analysis was performed using the two-tailed student’s t test, Fisher’s test and ANOVA. Results: The PI3K/Akt signaling pathway was observed to be highly active in the drug-resistant cells, and their level of PTEN was significanctly low compared with parental SKBR3 cells. Delivery of antisense PTEN duplex siRNA significantly decreased the trastuzumab chemosensitivity of parental SKBR3 cells, and marked activation of Akt signaling pathway was also recognized in oligonucleotid delivered parental cells. Moreover, in clinical analysis, immunohistochemical investigation revealed that trastuzumab treatment was remarkably successful in patients with elevated PTEN expression and statistically significant (p<0.05). Conclusions: PTEN activity might play an important and major role in its HER2/PI3K/Akt-mediated anti-tumor effect, and could be a useful biomarker for predicting the efficacy of trastuzumab in the treatment of breast cancer.

No significant financial relationships to disclose.
Oropharyngeal squamous cell cancers (OSCC) bearing transcriptionally active human papillomavirus (HPV) display distinct protein expression profile
Authors: Weinberger P. , Kountourakis P. , Sasaki C. , Haffty B. , Camp R. , Rimm D. , Psyrri A.
Published: March 2020

10028

Background: It is known that among HPV DNA+ OSCCs only the transcriptionally active ones are biologically and clinically relevant. pRb downregulation by HPVE7 results in p16 upregulation. We previously showed (JCO, Feb 2006) that p16 expression defines transcriptionally active HPV+ tumors. Thus, OSCC tumors can be Class I: HPV−/p16−, Class II: HPV+/p16−, or Class III: HPV+/p16+. We hypothesized that tumors classified by these criteria would show different protein expression profiles. Methods: Following institutional review board approval, paraffin-embedded specimens from 77 patients were collected from Yale-New Haven Hospital archives. Patients with OSCC treated with radiotherapy (RT) or surgery and RT were eligible. We used real-time polymerase chain reaction for HPV 16 and immunohistochemistry for p16 protein expression to classify 77 patients with OSCC. Tumors were classified into a 3-class model based on p16 expression and HPV DNA presence: Class I (HPV−, p16−) Class II (HPV+, p16−) and Class III (HPV+, p16+). A tissue microarray including these cases was constructed and examined for 13 target proteins with known involvement in tumor suppression or progression. We used a method of in-situ automated quantitative protein expression analysis (AQUA). Protein signal (AQUA score) was scored on a scale of 0–255.Protein expression between groups was analyzed by analysis of variance (ANOVA). Results: Thirty tumors were class I (39%), 29 were class II (38%), and 18 were class III (23%). There were significant differences for protein expression of Met (p=0.005), β-catenin (p=0.009), pRb (p=0.001), p53(p=0.026), epidermal growth factor receptor (EGFR) (p=0.009) and Vascular Endothelial Growth Factor (VEGF) (p=0.028). There was no difference for ki-67, COX2, p27, p21 or Cyclin D1 expression, while ERK2 and p14 trended towards significance (p=0.09 and 0.054, respectively). Conclusions: We demonstrated that tumors classified by HPV DNA presence and p16 expression status have different molecular phenotypes. Of particular note, class III tumors demonstrated increased expression of Met, β-catenin, EGFR and VEGF. These findings may prove useful for patient selection for molecular-targeted therapies.

[Table: see text]
Detection of novel variant TMPRSS2 /ERG fusion transcripts suggests independent genomic alterations may underlie origin of multi-centric prostate cancer
Authors: Yoshimoto M. , Joshua A. M. , Chilton-Macneill S. , Bayani J. , Prasad M. , Fleshner N. , Finelli A. , Evans A. , Sweet J. , Squire J. , Zielenska M.
Published: March 2020

10029

Background: Most of the early successes in identifying chromosomal translocations in neoplasias came from the study of hematological malignancies and sarcomas, with limited evidence that consistent genomic rearrangements were present in epithelial malignancies. Recently it was reported that ∼75% of prostate cancers carry a genomic rearrangement leading to fusion of the TMPRSS2 locus to either the ERG or ETV1 genes (both ETS transcription factors). In the fusion gene, the androgen-sensitive promoter elements of TMPRSS2 are thought to mediate over-expression of these ETS transcription factors. Over-activity of the ETS family of transcription factors has been suggested to be involved in the transition from pre-neoplasia to carcinoma as they regulate genes involved in processes such as adhesion, motility, invasion and angiogenesis. Methods: Using both RT-PCR and FISH with published primers and BACs respectively we analyzed 15 samples of prostatic carcinoma from radical prostatectomies and sequenced a subset of the TMPRSS2/ERG fusions. Results: We have found ERG-TMPRSS2 fusion transcripts in 6 samples and no ETV1-TMPRSS2 fusions. Of the 6 fusion tumours, 5 were Gleason 7 and 1 was Gleason 9. Tumour stages ranged from T2a-T3b. One sample with multi-centric carcinoma exhibited 2 distinct in-frame rearrangements generating novel TMPRSS2 /ERG fusion transcripts. Variant I TMPRSS2/ERG transcript was 430 bp and it led to fusion of exons 1 and 2 of the TMPRSS2 gene with exons 5 and 6 of the ERG gene. Variant II TMPRSS2/ERG fusion transcript was slightly smaller at 350 bp and it led to fusion of exon 1 of the TMPRSS2 gene to exons 5 and 6 of the ERG gene. These novel transcripts appear to be smaller than the published fusion proteins but preliminary analysis suggests that all known regulatory and functional protein domains are maintained. Conclusions: The demonstration of two new TMPRSS2/ERG variant fusion transcripts in prostate cancer deserves further study to evaluate their functional impact and prognostic and pathological importance. Moreover the presence of two distinct transcripts within a single multi-centric tumor provides genomic evidence that independent clonal neoplasms can arise synchronously in prostate cancer.

No significant financial relationships to disclose.
Comparison of genomic changes between tumors detected by screening mammography versus other methods
Authors: Becker T. E. , Ellsworth R. E. , Ellsworth D. L. , Deyarmin B. , Hooke J. A. , Shriver C. D.
Published: March 2020

10033

Background: The prognosis in patients with breast cancer detected by screening mammography has been found to be significantly better than those with similar-sized tumors detected by other methods (e.g. physical examination). (Joensuu, et al. 2004). Given the similar pathological characteristics, outcome differences may be explained by molecular differences within the tumors. To address this question, the relationship between genomic instability and methods of detection was assessed using allelic imbalance (AI) in similarly sized primary breast tumors from patients with breast cancer identified by screening mammography versus other methods. Methods: Patients with stage I breast cancer identified by screening mammography (n=50) versus other methods (n=50) were selected. Samples were matched between the two groups for clinical information. DNA was extracted from pure populations of tumor cells obtained by laser microdissection. Referent DNA was obtained from blood or disease-free skin samples from each patient. A panel of 52 microsatellite markers from 26 chromosomal regions commonly altered in breast cancer was used to assess AI, determined by the peak height ratio (≤0.35) between alleles from tumor and referent samples. Fisher’s exact test and Student’s t-tests were used to test for significant differences in levels of AI. Results: When clinical parameters such as age, ethnicity, menopause and status of tumor hormone receptors or HER2 were examined between tumor groups, no significant differences were detected. The overall frequency of AI between tumors detected by screening mammography (21.8%) versus other methods (22.3%) was similar (P=0.93). When patterns of AI were examined, none of the 26 chromosomal regions detected here showed a significant difference in AI events between the two groups. Conclusions: In this study, we have shown that global chromosomal differences are likely not the cause for the difference in tumor detection and prognosis between patients whose tumors were detected mammographically versus those whose were not. Prognostic factors known to correlate with disease aggressiveness and recurrence (e.g. hormone receptor or HER2 status) were also shown to not be associated with this difference in tumor detection among our patients.

No significant financial relationships to disclose.
Members of the glutathione (GSH) and ABC-transporter families are associated with clinical outcomes in patients with diffuse large B-cell lymphoma (DLCL)
Authors: Andreadis C. , Gimotty P. A. , Wahl P. , Hammond R. , Houldsworth J. , Schuster S. J. , Rebbeck T. R.
Published: March 2020

10035

Background: Chemotherapy only cures 60–70% of DLCL patients (pts). While genes in both the GSH and ABC-transporter families have been implicated in drug resistance in other malignancies, their role in DLCL remains controversial. Methods: This study includes 94 DLCL pts with tissue expression data obtained prior to anthracycline-based chemotherapy from 2 oligonucleotide microarray datasets (Affymetrix) with annotated clinical information. Patient-level expression was derived for 10 key genes utilizing probe set matching and levels were adjusted for a common probe set signal. Freedom-from-progression (FFP) was analyzed using an accelerated failure-time regression model, stratified by the international prognostic index (IPI). CART software was used to develop a classification tree for the 2-year disease-free rate. Results: Based on IPI, 48% of pts were low risk, 23% were low-intermediate (int) risk, 24% high-int risk, and 12% were high risk. After therapy, 30 pts (32%) had disease relapse or progression within 2 yrs and 58 pts (62%) were disease-free; 6 (6%) had shorter follow-up. Expression of multi-drug resistance 1 (MDR1; ABCB1) was favorably associated with FFP (β=1.89, p=0.004), with a 100-point change in the expression signal resulting in a near-doubling of the time to failure (time ratio (TR) = 2.2, 95%CI: 1.3–3.6). Expression of GSH S-transferase μ, classes 1/2/4 (GSTM124) was also favorably associated with FFP (β=0.14, p=0.001); a 1000-point signal change resulted in a near-doubling of the time to failure (TR=1.8, 1.3–2.4). Based on our tree, 2 risk groups were identified using IPI and 3 genes: ABCB1, GSTM124, and GSH cysteine ligase modifier (GCLM). Thus, 89% (78%-96%) pts in the low-risk group (N=56) were disease-free at 2 yrs vs. 25% (11%-43%) pts in the high-risk group (p<0.001). Conclusions: We observed a significant favorable association between expression of genes in the GSH and ABC-transporter families, and FFP in pts treated for DLCL. Moreover, we generated 2 risk groups based on the IPI and the expression of 3 key genes, which we plan to validate in an independent dataset. Our findings suggest an additional role for these genes that are classically associated with drug resistance.

No significant financial relationships to disclose.
Screening for ATM sequence alterations in African-American women diagnosed with breast cancer
Authors: Hirsch A. E. , Atencio D. P. , Rosenstein B. S.
Published: March 2020

10037

Background: Women who are heterozygous for mutations in the ATM gene, ATM carriers, are reported to have an increased risk of breast cancer compared with noncarriers of the mutation. There are few data on breast cancer susceptibility mutations in African-American women, particularly the non-BRCA mutations. We set out to determine whether there is evidence that ATM represents a breast cancer susceptibility gene in African-American women. Methods: One hundred thirty-two African-American women were screened for ATM sequence alterations. Thirty-seven (28%) were women with a histological diagnosis of breast cancer (cases) and 95 (72%) were age-matched women who were seen for a screening mammogram and had no evidence of cancer (controls). Genetic variants were identified using Denaturing High-Performance Liquid Chromatography (DHPLC). Results: Eighteen of 37 (49%) cases and 54/95 (57%) possessed at least one ATM variant (p =0.256). Missense variants were seen on 16/37 (43%) cases and 66/95 (69%) controls. In addition, 5/37 cases and 24/95 controls possessed multiple ATM sequence variants (p=0.107). The most common polymorphisms were: 378 T ->A, seen on 7/37 (19%) cases and 26/95 (27%) controls (p=0.219); 2685 A-> G, seen on 5/37 (11%) cases and 7/95 (6%) controls (p=0.217); and 1254 A-> G, seen on 2/37 (3%) cases and 9/95 (9%) controls (p=0.357). There were no significant differences in any of the single nucleotide polymorphisms (SNPs) detected between the cases and controls. Conclusions: We found no significant differences in the overall frequency of ATM variants between African-American women with breast cancer and an age-matched cohort of African-American women without breast cancer. ATM, therefore, does not appear to represent a breast cancer susceptibility gene in the African-American population. Many of the ATM variants that we found in high frequency were consistent with prior reports of common variants in patients of African ancestry. Comparing the difference in the frequency of ATM variants in the African-American population screened in this study with prior reports on non-African-American subjects will enable us to characterize the differential distribution of genetic alterations in various patient populations and lead to more optimally tailored cancer therapy.

No significant financial relationships to disclose.
Utility of clinical markers to identify patients with high risk cervical dysplasia and cancer
Authors: Ueda S. , Carney M. , Killeen J. , Wakabayashi M.
Published: March 2020

10055

Background: The natural history of cervical neoplasias appears unpredictable with 1/3 to 1/2 of CIN lesions regressing without intervention; therapeutic targeting aids in screening and selecting patients at risk and elucidating treatment response. This study aimed at determining if a pattern of tumor suppressor, oncogene, and growth factor expression (specifically HER-2/neu, progesterone and estrogen receptors (ER/PR), p53, and Ki67) exists in the progression of cervical cancer utilizing the widely applicable process of immunohistochemistry. Methods: Paraffin sections from 128 cervical biopsy, cone, and hysterectomy specimens collected from 1/00–12/04 were obtained from the surgical pathology repository at a tertiary care center with a primarily Asian and Polynesian patient population. Levels of expression in normal, low and high grade dysplasias, and invasive carcinomas were measured via automated immunohistochemical analysis. Results: 30% of samples came from patients who smoked. Ki-67 index rose with increasing dysplastic change (17–43%) but dropped with invasive carcinomas (<17%). HER-2/neu staining occurred at greater levels in high grade lesions and invasive carcinomas but average intensity measured <1, with >2.2 being of prognostic significance because of candidacy for Herceptin therapy by breast criteria. ER/PR expression generally remained low with a penchant for parabasal cells (2–14%), and p53 levels likewise appeared minimally affected (3–5%). Conclusions: While HPV infection immortalizes cervical cells via a p53/pRb pathway, the equivocal expression of wild-type p53 across pre-invasive and more aggressive lesions suggests the involvement of other environmental toxins and genetic mutations. PR and ER play only a minor role in the development of adenosquamous carcinomas, indicating a lack of hormonal control in cervical tumor growth and proliferation. Her-2/neu, however, participates in the late events of cervical malignancy but at such levels that clinical usefulness proves questionable. Although HER-2/neu perhaps holds limited prognostic value in cervical carcinogenesis, similar molecular pathways independent of HPV need to be investigated as adjuncts to the common Pap smear.

No significant financial relationships to disclose.
A lung cancer genomic risk prediction model derived from paraffin-embedded tissue
Authors: Harpole D. H. , Joshi M. M. , Petersen R. P. , Conlon D. H. , Tanaka K. , Shimizu D. , Kuramochi H. , Williams M. , Danenberg P. V. , Danenberg K.
Published: March 2020

10059

Background: We previously developed a validated fresh tissue-based genomic risk model in patients with early stage non-small cell lung cancer (NSCLC) using the Affymetrix U133 plus 2.0 Genechip. Limitations of this fresh tissue-based model include the need for immediate processing and limited availability; however, formalin-fixed, paraffin-embedded (FFPE) tissue is readily available and archived on every patient resected in North America. We investigated the ability of gene expression profiles generated on DNA microarrays using RNA isolated from FFPE NSCLC specimens to distinguish short-term and long-term survivors. Methods: Five to ten 5 um sections of FFPE tumor were collected from 61 NSCLC patients consisting of equal numbers of long- (+5-year) and short-term (<2 year cancer death) survivors. Fifty-five samples were microdissected (6 samples contained no tumor tissue) and RNA was extracted using a proprietary procedure of Response Genetics, Inc. For this feasibility study, Actin 300 < 30 cTs was chosen as a threshold for adequate RNA quantity for amplification to the GeneChip. Amplification and labeling of RNA were done using the Affymetrix two cycle amplification kit. The resulting cRNA was successfully hybridized to the U133 plus 2.0 GeneChip in 54/55 samples (98%). Data were analyzed using the SAM statistical software with Kaplan Meier survival analyses. Results: All analyses were performed using unsupervised hierarchical clustering and blinded duplicate samples had nearly identical gene expression profiles, indicating reproducibility. Adenocarcinoma segregated from squamous cell carcinoma with 98% accuracy (p=0.00004). A differentially expressed gene list between long and short survivors was determined. Distinct gene clusters were observed within each histological type segregating the tumors according to outcome. Kaplan Meier survival analysis stratifying on these clusters revealed significant differences in survival (cluster 1 and cluster 2 median survival>75 mos. vs. 30 mos., respectively; p<0.001). Conclusions: We have demonstrated the feasibility of creating a preliminary genomic risk prediction model using FFPE NSCLC tissue. Data will be presented on a larger training set (100+ patients) and a separate validation cohort of 100 patients.

[Table: see text]
Competing risk events determining probability of cause-specific failure in nasopharyngeal carcinoma
Authors: Huang Y. , Zee B. , Lam M. , Teo P.
Published: March 2020

10082

Background: The commonly employed Kaplan-Meier (KM) method is based on the assumption that different failure types (local-regional, distant, etc.) are independent. In reality, these failures occur at different stages in disease progression and are strongly correlated with each other. The assumption of independence of different failure types may violate certain assumptions in the modeling, and hence may affect the clinical interpretation and treatment selection. A better approach to estimate cause-specific failure probability is to calculate cumulative incidence rates by taking into account other events within a competing risk framework, in which the dependency of different failures are considered. Methods: The data was based on a large retrospective cohort study conducted at the Prince of Wales Hospital in Hong Kong, China, in 1996–97. 945 patients with nasopharyngeal carcinoma (NPC) had been treated with a standard protocol and been followed up regularly with a median follow-up period of 69 months (1–122 months). We calculated the cumulative incidence rates of local-regional failure and distant metastasis, and compared the result against KM method. In competing risk analysis, local regional failure, distant metastases and death were considered as competing events during the remaining lifetime of NPC patients from first presentation. Results: The probability of local-regional failure and distant metastasis was higher by KM method than by competing risk method. The result indicated that KM analysis overestimated event rate and the difference became larger in a longer follow-up period, when more competing events occurred. Conclusion: Kaplan-Meier analysis overestimates the probability of cause-specific failure. Competing risk analysis provides us a more accurate method in the determination of the pattern of failure. It provides better evidence to clinicians to enable them to predict the prognosis and select proper therapy.

[Table: see text]

No significant financial relationships to disclose.
Persistence of disseminated tumor cells (DTC) in bone marrow (BM) of breast cancer patients predicts increased risk for relapse—Results of pooled European data
Authors: Janni W. J. , Wiedswang G. , Fehm T. , Jueckstock J. , Borgen E. , Rack B. , Braun S. , Sommer H. , Pantel K. , Naume B.
Published: March 2020

10083

Background: The prognostic significance of DTC in the BM of breast cancer patients at the time of primary diagnosis has recently been confirmed by a large pooled analysis. If the persistence of DTC after adjuvant therapy confers a similar risk for relapse, there might be an indication for secondary adjuvant treatment. Methods: We analyzed BM aspirates of 697 patients from academic breast cancer units in Oslo (n=356), Munich (n=228) and Tuebingen (n=113) during recurrence-free follow-up at a median interval of 32.4 months (standard deviation [std] 19.4 mon) after primary diagnosis of breast cancer pT1–4, pN0–3 pM0. Carcinoma cells were detected using a standardized immunoassay with the monoclonal antibodies A45-B/B3 (Munich, Tuebingen), or AE1 and AE3 (Oslo), directed against cytokeratin (CK). Patients were followed for a median of 54.2 months (std 24.5 mon) after primary diagnosis. Results: Persistent DTC in the BM were detected in 15.6% of the patients (n=109). The Kaplan-Meier estimate for mean distant relapse-free survival estimate was 155.6 mon (142.4 - 168.9 95%CI) in patients with negative and 102.3 mon (93.6 - 111.0, 95% CI, p< .0001, log rank test) in patients with positive BM status. Patients without evidence of persistent DTC had a significantly longer overall survival (164.4 [155.6 - 173.3]), than patients with positive BM status (101.7 mon [89.4 - 113.9], p< .0001). In multivariate Cox regression analysis, allowing for bone marrow status, tumor size, nodal status, histopathological grading and hormone receptor status, DTC was of higher independent prognostic significance for subsequent reduced breast cancer specific survival (RR 5.9, 2.8 - 12.8, 95% CI, p< .0001), than nodal status at time of primary diagnosis (RR 1.2, 1.0 - 1.3, 95% CI, p=.014). Conclusion: Evidence of persistent DTC in breast cancer patients indicates an increased risk for subsequent relapse, and may serve for monitoring in future clinical trials. Such trials might investigate the benefit of individualized secondary adjuvant treatment or extended adjuvant therapy of patients with DTC.

No significant financial relationships to disclose.
Phase II breast cancer chemoprevention study with celecoxib in women at increased risk for breast cancer
Authors: Arun B. , Valero V. , Yin G. , Babiera G. , Murray J. L. , Browne D. , Gong Y. , Cook E. , Hortobagyi G. N. , Sneige N.
Published: March 2020

1010

Background: Short-term chemoprevention trials offer a convenient model to screen chemopreventive agents and identify endpoint biomarkers. One of the potential agents is celecoxib (C), which has antiproliferative and apoptosis inducing properties. In this prospective study, our primary aim was to evaluate changes in proliferation induced by C in breast tissue of high risk women. Here, we report changes in estrogen receptor (ER) proliferation index. Methods: 42 eligible high risk women were enrolled into the study, underwent fine needle aspiration (FNA) and started celecoxib treatment at 400 mg BID. Median age: 51.9 years. Risk factors: Gail risk > 1.67% (n=13), lobular carcinoma insitu (n=13), atypical hyperplasia (n=11), previous history of breast cancer (n=5). For ER and Ki-67 testing, thin preparations slides were incubated with primary mouse monoclonal antibody 6F11 against the ER and clone MIB-1, respectively. Appropriate negative and positive controls were included. At least 100 epithelial cells were evaluated per slide. Immunoreactivity for each marker was scored as the percentage of positive nuclei. We assessed the difference in ER and Ki-67 levels before and after treatment using a Wilcoxon signed rank test. Results: The average pre-treatment ER expression in FNA samples was 35.9% and Ki-67 was 2.4%. 19 (45%) showed hyperplasia or atypical hyperplasia. 39 patients underwent also post-treatment FNAs. The pre-and post treatment ER expression in this group was 35.7% (range 0–100%) and 27.4% (range: 0–100%), respectively. The difference in ER levels was statistically significant (p = 0.04). Twenty-six patients had Ki-67 levels measured both before and after treatment. The median difference in Ki-67 levels was 0 (range 0- 5). This change was not statistically significant (p = 0.63). Conclusions: We have completed accrual to a prospective short-term chemoprevention trial with celecoxib. We have found a significant downregulation of ER expression with 6 months celecoxib. Since ER expression is a marker of proliferation, this finding confirms celecoxibs antiproliferative properties. Currently, we have not observed a change in Ki-67; this could be partly due to the small number of samples and the fact that Ki-67 is low in normal epithelium.

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Selective spatial up-regulation of intratumoral stromal aromatase in breast cancer patients: Evidence for imbalance of local estrogen metabolism
Authors: Singer C. F. , Fink-Retter A. , Gschwantler-Kaulich D. , Hudelist G. , Czerwenka K. , Kubista E.
Published: March 2020

10105

Background: The suppression of local estrogens levels is of key importance in the treatment of ER positive breast cancer. Essentially all endocrine strategies act by either suppressing estrogen formation or by competitively inhibiting receptor binding in tumor cells. Nevertheless, little is still known about the local expression of aromatase and sultotransferase which are key modulators of intra-tumoral estrogen levels. Methods: We have performed immunohistochemostry to investigate the expression of aromatase and sulfotransferase in 42 samples obtained directly from malignant breast tumors, and compared it to biopsies obtained from uninvolved tissue in the vicinity of the invasion front, and to distant breast tissue. Results: We found that aromatase was equally detectable in both tumor epithelial and stroma, but was mostly epithelial in non-malignant tissues (p=0.00008, Fisher’s Exact Test). Also, aromatase protein expression was significantly more common in tumoral stroma when compared to peri-tumoral and distant breast stroma (p=0.00005, and p<0.00001, respectively). With the notable exception of cystosarcoma phylloides, sulfotransferase protein was only detectable in epithelial tissues, regardless of the location within the diseased breast. Epithelial sulfotransferase was, however, correlated with epithelial aromatase (r=0.35461, p=0.0009, Spearman’s Rho test) and with the epithelial ER status (r=0.29313, p=0.005). Conclusions: Taken together, we have demonstrated a differential aromatase and sulfotransferase protein expression pattern that is dependent of the spacial relation to a malignant breast tumor. Our results indicate a net increase in intra-tumoral active estrogen levels through increased stromal aromatization, while physiological local inactivation by sulfotransferase activity remains essentially unchanged.

No significant financial relationships to disclose.
Cervical cytologic screening: Optimal management of atypical squamous cells of undetermined significance in young women
Authors: Gibb R. K. , Wright J. D. , Davila R. , Powell M. A. , Rader J. S. , Gao F. , Mutch D. G.
Published: March 2020

1019

Background: HPV testing is a cost-effective strategy for the management of ASC-US cytology. However, young women have a high prevalence of transient HPV infections and low incidence high grade cervical lesions which may limit the utility of HPV testing in these women. We sought to determine the utility of HPV testing for young women with ASC-US. Methods: A review of all patients with ASC-US cytology between January, 2003 and December, 2004 was performed. Reflex HPV testing using the Hybrid Capture II system was performed on all samples. Age stratified rates of HPV positivity, rates of high grade dysplasia and sensitivity and specificity were estimated. Results: A total of 1290 women with ASC-US cytology were identified. The percentage of ASC-US samples that were HPV positive was highest in the young women and decreased with advancing age: < 25 yo (55%), 26–40 (36%), 41–50 (13%), > 50 yo (12%) (p<0.0001). Among HPV positive women with ASC-US, the identification of high grade lesions (HSIL, CIN2, CIN3, cancer) was lowest in the young women and increased with age: < 25 (12%), 26–40 (24%), 41–50 (19%), > 50 yo (24%) (p=0.05). A negative HPV test more reliably excluded high disease in the young women. No high grade lesions were detected in the HPV negative women ≤ 25 compared to 3.8% of those > 50 (p=0.04). The sensitivity for detection of high grade disease was higher in women < 25 compared to those > 50 (100% vs. 50%), while specificity was lower (14% vs. 44%). Conclusions: Given the high prevalence of HPV and low occurrence of high grade lesions in young women with ASC-US, an HPV based triage strategy will result in the referral of a large number of women for colposcopy and may limit its cost effectiveness. A program of repeat cytology may be more appropriate for the follow-up of young women with ASC-US.

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NSAIDs, osteoarthritis, and colorectal cancer risk: Can administrative data provide a window into the effectiveness of a cancer prevention?
Authors: Lamont E. B. , Dias L. E.
Published: March 2020

1022

Background: Randomized trials show that non-steroidal anti-inflammatory drugs (NSAIDs) reduce the risk of pre-cancerous polyps and may reduce the risk of invasive colorectal cancer (CRC), but studies of effectiveness have not been reported. Given the ubiquity of NSAIDS, existing population-level data sources may be leveraged to provide a window into the effectiveness of NSAIDs in CRC prevention. Methods: With data from the National Center for Health Statistics’ National Ambulatory Medical Care Survey, we established the suitability of the diagnosis of osteoarthritis (OA) as an instrument or marker of NSAID therapy among the elderly. With data from the National Cancer Institute’s SEER-Medicare program, we then identified elderly Medicare beneficiaries diagnosed with CRC in 1995 and compared them to elderly Medicare beneficiaries without histories of CRC according to antecedent OA diagnoses in Medicare claims files. Using logistic regression, we estimated the odds of CRC diagnosis among elderly Medicare beneficiaries with histories of OA compared to those without. As a control analysis, we also estimated the odds of a breast cancer diagnosis among those elderly female Medicare beneficiaries with histories of OA compared to those without. Results: Within the SEER-Medicare data files, we identified 4,599 elderly patients with a SEER diagnosis of invasive CRC and 100,270 patients without a SEER diagnosis of invasive CRC. Individuals with a diagnosis of OA in Medicare claims in the prior three years had an 18% lower risk of being diagnosed with CRC than individuals whose claims did not reflect antecedent OA (OR 0.82, 95% CI: 0.77–0.88). No association between antecedent OA and subsequent breast cancer (OR 0.99, 95% CI: 0.92–1.05) was found. Conclusions: In this case-control study that treats certain ICD-9 codes within Medicare claims as instruments of NSAID use, we find that elderly Medicare beneficiaries with ICD-9 codes for OA have an18% lower odds of being diagnosed with invasive CRC. Further, the magnitude of the CRC odds reduction is similar to what would be expected from the excess NSAID use alone.

No significant financial relationships to disclose.