bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression profiles can predict both general chemotherapy and gemcitabine-specific responses in breast cancer patients receiving neoadjuvant therapy
Authors: Oh D. S. , Carey L. A. , Fan C. , Sawyer L. , Hu Z. , Ma D. , Perou C.
Published: March 2020

10008

Background: The identification of markers predictive of neoadjuvant chemotherapy response would be an important advance. Therefore, we sought to identify gene expression profiles predictive of neoadjuvant response and evaluated their drug specificity. Methods: DNA microarray analysis was performed on pre-treatment core biopsies from two different locally advanced breast cancer patient sets receiving neoadjuvant chemotherapy; the first set (L9819) was treated with 4 cycles of doxorubicin + cyclophosphamide (AC) followed by 4 cycles of paclitaxel (T) or paclitaxel + trastuzumab (TH). The second patient set (S329) received 4 cycles of gemcitabine + doxorubicin followed by 4 cycles of gemcitabine + cisplatin. Clinical response was based on RECIST criteria. Results: We obtained successful microarrays on 44 (L9819) and 46 (S329) of the patients enrolled. Ten-fold cross-validated supervised analyses separately performed on each dataset identified gene expression patterns that accurately predicted (75–85% accuracy) response (clinical complete responders vs. others) evaluated on those patients that completed all 8 cycles of chemotherapy. The predictive expression patterns from the L9819 trial were significantly enriched for apoptosis genes, while the predictive gene set from the S329 trial was enriched for nucleotide metabolism genes. The L9819 predictor was able to accurately predict (75% accuracy) response for the S329 patients, however, the S329 predictor was not able to predict response on the L9819 trial (50% accuracy). We speculate that the L9819 predictor was able to predict response on the S329 dataset (despite the fact that only doxorubicin was common to both trials) because this profile was enriched for apoptosis genes that may reflect a general mechanism of chemotherapy response/resistance, while the S329 predictor was more tuned to gemcitabine’s specific mechanism of action. Conclusions: This study may provide the means to predict response to AC-T(H) and GA-GC neoadjuvant chemotherapy. The ability of one predictor to work on the other dataset, but not vise versa, suggests that both general chemotherapy and drug specific response profiles may have been identified.

[Table: see text]
Prostate cancer screening, treatment, and outcomes— A population-based study
Authors: Lu-Yao G. L. , Albertsen P. C. , Stanford J. L. , Stukel T. A. , Walker-Corkery E. S. , Barry M. J.
Published: March 2020

10017

Background: It remains unknown whether more intense prostate cancer screening and treatment reduces prostate cancer mortality. We describe the experiences of two population-based cohorts with 15-years follow-up (1987–2001) to address the following questions: 1) does more intense screening and treatment for prostate cancer lead to lower mortality in community settings, and 2) do older men (age ≥75–79) benefit from more intense prostate cancer screening and treatment. Methods: Population-based cohort study consisting of white male Medicare beneficiaries who resided in the regions covered by the Seattle (N=88,863) and Connecticut (N=114,785) cancer registries. Inclusion criteria are age 65–79 and free of prostate cancer on January 1, 1987. All study subjects were followed through death or the end of 2001. The main outcomes are rates of screening for prostate cancer, treatment with radical prostatectomy, external beam radiotherapy, and prostate cancer specific mortality. Results: Between 1987 and 1990, compared to men in Connecticut, men in the Seattle region were 5.4 times (95% C.I. 4.8 - 6.1) more likely to undergo PSA testing, 2.2 times (95% C.I. 1.8 - 2.7) more likely to under go prostate biopsy, 5.9 times (95% C.I. 5.5 - 6.9) more likely to have radial prostatectomy, and 2.3 times (95% C.I. 2.2 - 2.5) more likely to have external beam radiation. The cumulative risk of radical prostatectomy or external beam radiation reached 9.1% in the Seattle cohort and 5.0% in the Connecticut cohort in 2001. After 15 years of follow-up, prostate cancer mortality rates were similar for subjects in the two study regions (hazard ratio of Seattle to CT: 1.01, 95% C.I. 0.93 - 1.09). For older men (aged 75–79 in 1987), however, the prostate cancer mortality rate was slightly higher in the Seattle than the Connecticut cohort (hazard ratio: 1.16, 95% C.I. 1.02 - 1.32). Conclusion: More intense screening for prostate cancer, surgery or radiation among a cohort of Medicare beneficiaries in the Seattle area compared with their counterparts in Connecticut has not lead to significantly lower mortality from prostate cancer over 15 years of follow-up.

No significant financial relationships to disclose.
A novel role of DNA fragmentation factor as a tumor suppressor through maintaining genomic stability
Authors: Yan B. , Wang H. , Bedford J. S. , Dewhirst M. W. , Li C.
Published: March 2020

10023

Background: DNA fragmentation is a hallmark of apoptosis. However, the biological function of apoptotic DNA fragmentation remains unclear. Genes encoding the nuclease responsible for DNA fragmentation, DFF40/CAD and DFF45/ICAD, are deleted, mutated, or aberrantly expressed in many human tumor types. Abnormalities in this gene are associated with aggressive tumors and poor prognosis in cancer patients. Tumor-specific DFF45 gene mutations were identified in human tumors, indicating the involvement of this gene in tumor development. Methods: We studied genetic instability, cellular transformation and tumorigenesis in cells and mice deficient in DNA fragmentation. We applied two methods to inhibit DNA fragmentation, overexpressing a modified ICAD which inhibits DNA fragmentation during apoptosis in stable cell lines and disrupting cad gene in transgenic mice. Genetic instability was studied by gene amplification assay, mutation assay and cytogenetic analysis including chromosomal aberrations and translocations. Cellular transformation was studied by soft agar assay using CAD-/- MEF cells. Tumorigenesis in DFF/CAD-knockout mice was studied by radiation carcinogenesis and two-stage skin chemical carcinogenesis. Results: (1) Human and mouse cancer cell lines with their CAD activity inhibited exhibited significantly more genetic instability under spontaneous or mutagen (ionizing radiation) induced conditions. These are reflected as elevated frequencies of chromosomal aberrations, gene amplifications, and gene mutations; (2) Mouse cells with targeted disruption of the cad gene exhibit a similar, increased level of genetic instability when the host animal is irradiated; (3) Mechanistically, CAD maintains genetic instability through the removal of cells with DNA damage, a role that is similar to genomic “gatekeepers” like p53; (4) CAD-null mouse embryo fibroblasts exhibited enhanced cellular transformation; (5) Significantly enhanced susceptibility to chemical and radiation-induced tumor development was observed in mice with targeted disruption of the CAD gene, indicating that it plays a role in suppressing tumor development. Conclusions: CAD plays an important role in maitaining genetic stability and suppressing tumor development.

No significant financial relationships to disclose.
EGFR mutations and human papillomavirus in squamous cell carcinoma of tongue and tonsil
Authors: Na I. , Kang H. , Park Y. , Koh J. , Cho S. , Lee B. , Lee Y. , Shim Y. , Ryoo B. , Yang S.
Published: March 2020

10027

Background: Some studies reported the presence of somatic mutation of EGFR in head and neck cancer. Human papillomavirus (HPV) was associated with some types of head and neck cancer. We performed this study to determine clinical significance of EGFR mutation and its association with HPV infection in patients with tongue and tonsil cancer. Methods: We sequenced exons 18–21 of EGFR TK domain and HPV typing was performed using the HPV DNA chip in tissues of patients with tongue and tonsil cancer. Univariate and multivariate analysis was performed. Results: Eighty-one patients were included. We detected EGFR mutation in 10 (12%) patients. Mutation was significantly frequent in never-smoker (21% versus 5%; P = 0.031). The mutation frequency did not differ between tongue and tonsil cancer (12% vs. 13%, respectively; P = 0.978). There was no difference between female and male (12% vs. 14%, respectively; P = 0.754). Eleven of 81 (14%) patients were HPV positive. HPV positivity was more common in young (<60 years) than older (≥60 years) patients (20% vs. 3%, respectively; P = 0.039). Patients with tonsil cancer showed more frequent HPV positivity than tongue cancer (42% vs. 2%, respectively; P < 0.001). There was no association between HPV positivity and EGFR mutation. In terms of disease-free survival, patients with EGFR mutation had better survival independently of stage and primary site (hazard ratio, 0.246; 95% CI, 0.075 to 0.802). Conclusions: We did not find association between EGFR mutation and HPV positivity. Patients with EGFR mutation showed better outcome in tongue and tonsil cancer. These results suggest that EGFR mutation may be a prognostic factor in patients with head and neck cancer.

No significant financial relationships to disclose.
Multi-center prospective analysis of risk-reducing salpingo-oophorectomy to prevent BRCA-associated breast and ovarian cancer
Authors: Kauff N. D. , Domchek S. M. , Friebel T. M. , Lee J. B. , Roth R. , Robson M. E. , Barakat R. R. , Norton L. , Offit K. , Rebbeck T. R.
Published: March 2020

1003

Background: Our groups previously reported on the efficacy of risk-reducing salpingo-oophorectomy (RRSO) for the prevention of BRCA-associated breast and ovarian cancer. (Kauff ND, et al. NEJM 2002; Rebbeck TR, et al. NEJM 2002) Limitations of those reports included relatively short prospective follow-up and lack of power to analyze the protection of RRSO when participants were stratified by BRCA1 vs. BRCA2. To address these limitations, we have pooled our updated datasets to provide robust estimates of the efficacy of RRSO. Methods: 886 women ≥ 30 years of age, with a deleterious mutation in BRCA1 or BRCA2 and ovaries in-situ at time of genetic test results, were enrolled on prospective follow-up studies at one of eleven centers from 11/1/1994 - 12/1/2004. Women chose to participate in either ovarian surveillance or undergo RRSO. Follow-up information was collected by questionnaire and medical record review. Follow-up time was counted from time of RRSO or from time of genetic test results for women who did not undergo RRSO. After excluding cancers diagnosed within the first 6 months of follow-up, the effect of RRSO on time to diagnosis of breast or BRCA-associated gynecologic cancer was analyzed using a Cox proportional-hazards model. Results: 559 (63%) participants underwent RRSO a median of 5 months after genetic test results. 12 occult ovarian or fallopian tube cancers were diagnosed at time of RRSO. During a mean 40 months follow-up, RRSO was associated with a 52% reduction in breast cancer risk and a 91% reduction in ovarian cancer risk (see Table ). When the cohort was stratified by mutation status, RRSO was associated with a reduced risk of BRCA1-associated ovarian cancer and BRCA2-associated breast cancer. Conclusions: The results confirm that RRSO is highly protective against BRCA-associated breast and ovarian cancer. These results also generate the hypothesis that the protection conferred by RRSO against specific cancers may differ between carriers of BRCA1 and BRCA2 mutations.

[Table: see text]

No significant financial relationships to disclose.
Promoter methylation and protein expression of the Slit2 and Robo1 genes in resected esophageal squamous cell carcinoma
Authors: Tzao C. , Hsu H. , Lee S. , Hsu W. , Sun G. , Cheng A. , Wang Y.
Published: March 2020

10032

Background: Recent epigenetic studies suggest that the slit-robo pathway is involved in tumorigenesis of a number of human cancers and a role of SLIT2 as a tumor suppressor gene. In view of lack of epigenetic studies of SLIT2 and ROBO1 genes in squamous cell esophageal carcinoma (ESCC), we investigate whether promoter methylation is responsible for aberrant protien expression of SLIT2 and ROBO1 and whether promoter methylation and aberrant protein expression of these two genes are related to prognosis after resection for ESCC. Methods: We analyzed promoter methylation of SLIT2 and ROBO1 using methylation-specific polymerase chain reaction (MSP) and their protein expression using immunohistochemistry in 60 resected tumor specimens. The Pearson χ2 test was used to compare promoter methylation and protein expression of SLIT2 and ROBO1 among patients with different clinicopathological parameters. Concordance analysis was performed between promoter methylation and protein expression of these two genes. Results: Promoter methylation of SLIT2 and ROBO1 was observed in 43 (71.6%) and 16 (26.6%) of the 60 resected specimens, respectively (P = 0.002). Aberrant protein expression of SLIT2 and ROBO1 was observed in 51 (85%) and 23 (38.3%), respectively (P < 0.001). Promoter methylation of SLIT2 correlated well with smoking (P = 0.046), betel nut chewing (P = 0.032), staging (P = 0.018) and tumor (T) factor (P < 0.001) of the TNM system and its aberrant protein expression correlated well with betel nut chewing (P = 0.038), staging (P = 0.025) and nodal (N) involvement (P = 0.004). Promoter methylation and protein expression of ROBO1 did not correlate with any of these clinicopathologic parameters. A concordance rate of 88.2% and 74.2% was present between expression of SLIT2 and ROBO1 protein and their promoter methylation (P < 0.001) and (P = 0.004), respectively. Conclusions: Aberrant expression of SLIT2 and ROBO1 protein is frequently associated with the presence of ESCC, with SLIT2 a more predominant one. Promoter methylation is one of the major mechanisms responsible for aberrant protein expression of these two genes in resected ESCC. Promoter methylation and aberrant protein expression of SLIT2 may predict prognosis after resection for ESCC.

No significant financial relationships to disclose.
Genetic models for estimating age-specific risk of sporadic breast cancer
Authors: Jupe E. R. , Ralph D. A. , Aston C. E. , Manjeshwar S. , Pugh T. D. , Gramling B. A. , Defreese D. C. , Shimasaki C. D.
Published: March 2020

10038

Introduction: Accurately assessing individual risk of developing breast cancer is essential for early detection and prevention. The possibility exists to improve upon the widely used Gail model in predicting an individual’s risk of developing breast cancer, by incorporating both genetic and lifestyle information. Methods: We have developed a model with data from ∼7,000 Caucasian women (2,300 cases and 4,700 controls) enrolled in four geographically distinct regions of the US. Questionnaire-based information on clinical and lifestyle variables, including the Gail model questions, were collected. All samples were genotyped for 120 common, functional polymorphisms in candidate genes likely to influence breast carcinogenesis. The data were randomly divided into training (75%) and validation (25%) sets. Because of age-dependent genetic penetrance observed in the study, separate models were built for three age intervals (30–44, 45–54 and ≥ 55). Following assessment of Hardy-Weinberg equilibrium and linkage-disequilibrium, univariate and multivariate statistical analyses were performed to assess genetic main effects and epistatic interactions and define model terms. Models were evaluated by Receiver Operator Curve (ROC) analysis. Results: The riskprediction models developed for all three age intervals were informative with Area Under the Curve (AUC) that was consistently greater than expected by chance and exceeded the predictive power of the Gail model alone. No strong associations were found between scores derived from our models and the Gail model. The results suggest that the genetic markers examined contribute to breast cancer prediction independent of the Gail scores, and due to the improvement in the AUC, probably complement the Gail scores. The strongest genetic contributors to risk were identified in the youngest group where the AUC was 0.71 in the training set and 0.66 in the validation set. Estimated AUC values appear to be quite stable across 1,000 bootstrap samples of training and validation sets. Conclusions: The predictive power of these models integrating both genetic and epidemiological data provides an improved estimate of an individual’s breast cancer risk.

[Table: see text]
Phase II trial of iboctadekin (rhIL-18) on a daily X 5 schedule in metastatic melanoma (MM)
Authors: Kirkwood J. M. , Kefford R. , Logan T. , Mainwaring P. N. , Millward M. , Pavlick A. C. , Dar M. M. , Kathman S. , Laubscher K. , Bell W.
Published: March 2020

10043

Background: Interleukin-18 (IL-18) is an immunostimulatory cytokine with potent antitumor activity in preclinical models. Two phase I studies of recombinant human (rh) IL-18 explored a wide dose range (0.03–1.0 mg/kg) without reaching a maximum tolerated dose (MTD) on the daily × 5 schedule. Pharmacodynamic data including inflammatory cytokine production and activation of lymphocyte subsets revealed optimal biologic activity at the lower end of the dose range (0.01–0.2 mg/kg) as did 2 unconfirmed partial responses (PRs) in a MM and a renal cancer patient (pt) at 0.1 mg/kg. Methods: An open-label, randomized, phase II trial in 60 adult pts with previously untreated MM was conducted to evaluate the efficacy and safety of rhIL-18 administered as a 2-hour IV infusion daily × 5 every 28 days for 6 cycles. Pts with PS ≤ 1, without known CNS involvement, and with adequate end organ function were randomized in stage 1 to 3 dose levels of IL-18 stratified according to AJCC M stage 1a/b vs. 1c. Two confirmed responses for a given dose level in Stage 1 were required to enroll 20 additional pts/level in Stage 2. The 1° objective was determination of overall response rate (ORR) for each dose level. Progression-free survival (PFS), tolerability, and immunogenicity were 2° endpoints. Results: 64 pts were treated at 3 dose levels. Nine pts remain on study. One pt experienced a confirmed PR. Based on preliminary data, the difference in PFS 6 months (mos) was significant (p=0.03) for 0.01 vs 0.1 mg/kg. Most common toxicities were mild to moderate fever, rigors, chills, n/v, and headache. Anti-IL18 antibody (Ab) development correlated with dose level. No clinically significant adverse events were associated with Ab development. Conclusion: Iboctadekin has an acceptable tolerability profile and has activity in MM but insufficient confirmed responses have been observed at this time to initiate Stage 2. Preliminary PFS 6 months indicates an advantage for pts treated at the lowest dose.

[Table: see text]

[Table: see text]
Gene expression subtype and p53 mutational status are correlated among neoadjuvantly treated breast cancers in UNC LCCC9819 and I-SPY1 (CALGB 150007/ACRIN 6657)
Authors: Carey L. A. , Oh D. , Sawyer L. , Edmiston S. , Tolbert D. , Moore D. , Buxton M. , Esserman L. , Perou C. , Conway K.
Published: March 2020

10048

Background: LCCC9819 and I-SPY1 are designed to examine multiple potential predictive markers and assays simultaneously in biopsies from neoadjuvantly treated patients. Both p53 mutation status and molecular ‘intrinsic‘ subtype have been associated with chemosensitivity. In this preliminary analysis, we present the p53 mutation spectra by molecular subtype from patients enrolled in the combined LCCC 9819 and I-SPY1 datasets. Methods: Patients received neoadjuvant anthracycline-based chemotherapy with pre-therapy biopsies. p53 mutation analysis was performed by p53 GeneChip assay for point mutations/1 bp deletions followed by SSCP if GeneChip-negative. Abnormal GeneChip or SSCP were confirmed by sequencing. DNA microarrays used Agilent human microarrays and a common reference strategy approach. Results: 51 patients were treated on LCCC9819 and have available molecular analyses; 87 from I-SPY1. 111 patients have completed p53 mutation analysis, revealing a high proportion (42%) with abnormal p53. Of these, several were in known hotspots, including R175H (4 mutations), R248L (1), R273H (3), and R273C (2). Most were missense mutations (81%), however, there were 8 (17%) null and 1 (2%) silent mutations. Most (91%) mutations were in the DNA binding domain. Molecular subtypes are known on 111 tumors: 22% Luminal A, 22% Luminal B, 3% Normal-like, 23% HER2+/ER−, 27% Basal-like, and 3% unclassified. Among those with both assays complete, we found that p53 mutations differed by subtype (Table). Conclusions: P53 mutations are common in these independent neoadjuvant datasets, and differ in frequency between molecular subtypes, which may have implications for predictive factor identification. Future analyses will include aCGH, cell cycle protein analysis, proteomics, and clinical data including response to chemotherapy. (supported by UNC Breast SPORE-CA58223, NIH M01RR00046, DAMD 17–02–1-0521).

[Table: see text]

No significant financial relationships to disclose.
Identification of cancer of unknown primary with gene expression profiling
Authors: Li H. , Qu K. , Tokoro K. , Ren Y. , Liu J. Y. , Sferruzza A. , Bender R.
Published: March 2020

10052

Background: Patients with metastatic cancer of unknown primary (CUP) generally have a poor prognosis, with a median survival of 2–10 months. Conventional diagnostic approaches for identifying the primary tumor site are successful in only 20%-30% of cases; however, such identification provides prognostic information and helps with selection of tumor-specific therapy, leading to improved survival. Recent studies indicate that gene expression-based classification of CUP is highly successful in predicting the site of origin. We report herein development and validation of a method that determines the site of tumor origin by comparing the gene expression profiles of CUP cases to those in a database created from known tumor types. Methods: RNA extracted from frozen and formalin-fixed, paraffin-embedded (FFPE) tissue wasis purified and amplified using the Paradise Reagent System System (Arcturus, Mountain View, CA). Following reverse-transcription, cDNA products wereare used in a semi-quantitative real-time PCR to detect 87 tumor-associated genes and 5 reference genes in an ABI PRISM 7900HT Detection System (Applied Biosystems, Foster City, CA). Gene expression data wereare then compared to those in a database, composed of gene expression profiles of 571 samples from 39 different tumor types, using k-nearest neighbor analysis to predict the most likely site of tumor origin. Intra- and interassay reproducibility was determined. Frozen and FFPE tissues (n=57) from a well-characterized, independent sample set were also tested in a blinded manner to further validate the method. Results: Based on the real-time PCR cycle threshold, the intra- and interassay reproducibility ranged from 0.1%-4.3% and 0.5%-8.2%, respectively. The primary tumor type was identified in 77% of cases. The assay determined the correct tumor type in 88% (44/50) of the samples. Seven samples were not reported: 3 failed to amplify adequately and 4 had an unacceptably low confidence level. Conclusions: We have shown that gene expression profiling can determine the most likely site of tumor origin. Our data suggest that this new method is able tomay identify the primary site of tumor origin in 77% of CUP cases.

No significant financial relationships to disclose.
Chemokine receptor 5 (CCR5) Δ32 base pair somatic gene deletions from PBM predict survival of patients with advanced stage (IV) melanoma
Authors: Essner R. , Lee J. H. , Itakura H. , Huynh Y. , Scheri R. , Morton D.
Published: March 2020

10056

Background: Chemokines are a family of secreted chemotactic cytokines that function in leukocyte trafficking and activation. Chemokine receptor 5 (CCR5) is involved in CD4+ T-cell mediated immunity and dendritic cell (DC) activation. A 32 base pair (bp) deletion mutation on chromosome 3p21 result in a non-functional CCR5 with defective immunity. We hypothesized that CCR5Δ32 bp mutations would result in a diminished immune response to melanoma and maybe predictive of outcome of patients with advanced-stage disease. Methods: We identified 160 AJCC stage IV melanoma patients from our database whom had been made NED (no evidence of disease) by surgical resection. 105 (66%) had received active specific immunotherapy in phase I/II trials. Genomic DNA was extracted from peripheral blood mononuclear cells (PBM) and screened by PCR for the presence of CCR5Δ32 bp deletions. PCR products were sequenced to verify the result. Statistical analysis was performed by using Log-rank, Chi-square & Student’s t-test. Results: Of 160 patients, 100 (63%) were men. Ages ranged from 19–82 (median 47 yrs.). 46 (29%) patients had M1a, 48 (30%) Mlb and 66 (41%) Mlc stage of metastases prior to surgical resection. Median follow-up was 47.5 mos. 135 (84%) patients had wild-type (Wt)CCR5 gene; 21 (13%) had heterozygous (HtM) and 4(3%) had homozygous mutations (HoM). 5-yr survival was significantly higher for patients with CCR5 Wt than those with HtM or HoM deletions (55.6%±6.1% vs. 21.4%±10.8%, p=0.031). None of the 21 patients with HtM survived 10yrs. Multivariate analyses demonstrated only CCR5 gene status (Wt vs. HtM/HoM) as predictive of survival (p=0.007). Age (p=0.23), gender (p=0.38), and M stage (M1a vs. M1b/c) (p=0.31) were not important in the statistical model. When immunotherapy responses (+ vs. -) were included in the model both responses and CCR5 mutation status were significant (p=0.0021, p=0.0063 respectively). Conclusions: Our results suggest CCR5Δ32bp somatic mutations from PBM are present in advanced-stage melanoma patients and are highly predictive of survival. Functional CCR5 may be used as a selection factor for surgery but more importantly essential for either active immunotherapy or the natural immune response to be effective for these patients.

No significant financial relationships to disclose.
The adenocarcinoma-specific stage shift: Closely-repeated low-dose helical CT screening in a high-risk cohort for 10 years
Authors: Seki N. , Eguchi K. , Kaneko M. , Ohmatsu H. , Kakinuma R. , Matsui E. , Kusumoto M. , Tsuchida T. , Nishiyama H. , Moriyama N.
Published: March 2020

1006

Background: Trials of CT screening for lung cancer have not yet proven the presence of a stage shift, to say nothing of a mortality benefit. On the basis of an update from Anti-Lung Cancer Association project, we investigated whether the stage shift did occur during the long-term closely-repeated low-dose helical CT screening in a high-risk cohort. Methods: A total of2,120 participants (mean age 63 yrs, 87% male, and 83% smoker) underwent low-dose helical CT screening semiannually from 1993 to 2004 (median, 3.5 years). Results: Nineteen prevalence and 57 incidence lung cancers were detected. In comparison between baseline and repeated screening, the detection rate was 0.90% vs. 0.32% (P < 0.001) and the tumor size was 24mm vs. 17mm (P = 0.018). Adenocarcinoma (74% and 63%) and stage IA (58% and 79%) were observed most commonly in both screen-groups, respectively, but their proportions showed no significant difference between groups, respectively. Regarding the survival of lung cancer patients, screening type was not a significant prognostic factor. In repeated screening,the detection rate oflung cancer except bronchioloalveolar carcinoma increased significantly depending on CT repeating times (r = 0.50, P = 0.020). Moreover, the proportion of stage II-IV and the tumor size decreased significantly only in invasive adenocarcinoma (r = -0.77, P = 0.007 and r = -0.60, P = 0.029, respectively). Conclusions: The adenocarcinoma-specific stage shift did occur in a high-risk cohort. Considering larger proportion of adenocarcinoma in this population, smokers might benefit from the long-term closely-repeated low-dose helical CT screening.

No significant financial relationships to disclose.
Detection of mutations in the EGFR tyrosine kinase domain by DNA sequencing
Authors: Da Silva M. , Flynn K. E. , Choppa P. , Zhou Z. , Bell D. W. , Iafrate A. J. , Paler R. J. , Allitto B. A. , Rohlfs E. M.
Published: March 2020

10060

Background: The presence of sequence alterations in the tyrosine kinase domain of the epidermal growth factor receptor gene (EGFR) have been documented as gefitinib/erlotinib sensitizing mutations and serve as indicators for targeted (Iressa/Tarceva) therapy in patients with non-small-cell lung carcinoma (NSCLC). We report results obtained from the development and validation of a sequencing-based assay for the detection of these somatic mutations. Methods: Specimens tested included4 DNA samples with known EGFR mutations and 59 formalin-fixed primary or metastatic lung carcinomas, 22 which had known EGFR genotypes and 37 which were untested. Forty-one were adenocarcinomas. The most tumor-rich area of the section was micro-dissected to minimize amplification of non-tumor cells. DNA was extracted following treatment to deparaffinize and dehydrate the tissue section. A quantitative EGFR PCR was used to determine if the quantity of isolated DNA was sufficient for sequence analysis. A first-round multiplex amplification was followed by four semi-nested PCR amplifications targeting EGFR exons 18–21 and the flanking splice sites. These products were then subjected to sequencing reactions and evaluated for alterations as compared to the wild type sequence. Results: All of the specimens met the minimum quantity of 750pg of EGFR DNA required for the analysis. Thirty-one of the 37 anonymous specimens showed no sequence alterations in exons 18–21. Four of the five specimens with sequence alterations had known sensitizing mutations; three were L858R and one was 2235–2249del15. One specimen had a novel W817X mutation in exon 20. One specimen failed analysis for three of the exons. The percent tumor in the area selected for analysis varied from 5–30% in the specimens with sequence alterations. Twenty-five of the 26 samples with known genotypes were correctly identified by this method. In one sample a reported mutation was not detected, possibly due to selection of a different tumor nodule for testing. Resequencing of the sample by an outside lab confirmed the negative result. Conclusions: We have developed a highly sensitive and accurate sequencing-based assay capable of detecting somatic mutations in EGFR exons 18–21 which is important in the treatment and management of patients with NSCLC.

[Table: see text]
Therapeutic efficacy of an oncolytic adenovirus monitored by in vivo imaging of an ovarian cancer model
Authors: Tsuruta Y. , Pereboeva L. , Rein D. T. , Breidenbach M. , Curiel D. T.
Published: March 2020

10061

Background: Although a number of advances in ovarian cancer treatment have occurred in the last decade, most patients will experience a recurrence after standard therapies. Recently, virotherapy has been proposed as a new therapeutic approach for ovarian cancer. Conditionally replicative adenovirus (CRAd) contains tumor-specific promoters that restrict virus replication to cancer cells and has shown particular promise as oncolytic viral agents. However, the lack of a tumor-volume monitoring system hinders the evaluation of CRAd impact on cancer treatment. Therefore, methods for analyzing CRAd efficacy and tumor response are required. Mesothelin, a cell surface glycoprotein, is overexpressed in ovarian cancer but not in normal ovarian tissues. The purpose of this study was to explore the therapeutic utility of a mesothelin promoter-based CRAd in a murine model of ovarian cancer, using a non-invasive biological imaging system. Methods: We constructed a mesothelin promoter based CRAd which also contains a modified fiber (Ad5/3 fiber) previously shown to improve infectivity of many ovarian cancer cells (Ad5/3MSLN). Viral replication and oncolysis were assessed in a panel of ovarian cancer cell lines. To test the oncolytic efficacy of Ad5/3MSLN in murine model, firefly luciferase-expressing SK-OV-3-luc cells were injected intraperitoneally (i.p.), followed by an i.p. injection of viruses. Then, bioluminescence imaging of tumor luciferase activity was carried out. Results: Ad5/3MSLN achieved up to 10,000-fold higher cell killing effect and up to 120-fold higher levels of viral replication in all ovarian cancer cell lines tested, compared to wild type Ad5. In vivo tumor imaging confirmed that Ad5/3MSLN significantly inhibited tumor growth, while the untreated mice had rapid tumor growth (p<0.05). Survival with Ad5/3MSLN was significantly enhanced when compared with no virus, or wild type Ad5-treated group (p<0.05). Conclusions: The robust replication, oncolysis, and in vivo therapeutic efficacy of Ad5/3MSLN demonstrated that this CRAd is a promising candidate for treating ovarian cancer. Importantly, we have established an in vivo non-invasive imaging system, which has allowed repeated and longitudinal measurements of tumor growth after CRAd treatment.

No significant financial relationships to disclose.
Nuclear localization of NF-kappaB p65 in primary prostate tumors is predictive of pelvic lymph node metastases
Authors: Lessard L. , Bellon-Gagnon P. , Alam-Fahmy M. , Karakiewicz P. , Mes-Masson A. , Saad F.
Published: March 2020

10077

Background: Pelvic lymph node metastases are associated with a greater risk of prostate cancer recurrence and peripheral metastasis. Unfortunately, markers predictive of lymph node metastasis and/or recurrence after radical prostatectomy are limited and new molecular markers are needed to identify patients at higher risk of progression. NF-kB (p65) is a candidate molecular marker already associated with poor clinical outcomes such as biochemical recurrence and bone metastasis. We have also reported elevated nuclear p65 expression in prostate cancer lymph node metastasis. Pertinent to this issue, we tested whether the nuclear localization of p65 in radical prostatectomy specimens could predict the presence of lymph node metastases. Methods: Following informed consent, 51 patients who underwent radical prostatectomy were included in the study: 20 patients had lymph node metastasis at surgery and 31 patients had no evidence of lymph node metastasis and were used as the control group. All cases in the control group had no biochemical relapse 5 years following radical prostatectomy. NF-kB expression in prostate tumor sections was assessed by immunohistochemistry using a monoclonal NF-kB p65 antibody. The relation between nuclear p65 expression in primary tumors and lymph node metastasis was tested in univariate and multivariate Cox regression models. Results: Primary tumors of metastatic patients had an average of 21.25% of tumor cells with nuclear p65 expression as opposed to 9.42% of tumor cells of control patients (p=0.001). Univariate Cox regression demonstrated a 7.5% increased risk of having lymph node metastases for each percent increase in p65 nuclear staining (p=0.003). In the multivariate model, after controlling for pre-operative PSA (p=0.175), Gleason patterns (p=0.382), pathological stage (p=0.436), extracapsular extension (p=0.243) and seminal vesicle invasion (p=0.016), nuclear p65 was associated with an 8.8% increased risk for lymph node metastases (p=0.024). Conclusion: In univariate and multivariate analyses, p65 nuclear expression was strongly predictive of lymph node invasion. We propose that nuclear NF-kB (p65) may serve as a useful independent molecular marker for stratifying patients at risk for lymph node metastases.

No significant financial relationships to disclose.
Developing a predictive model for disease status in breast cancer patients using Th1 and Th2 serum cytokine profiles
Authors: Khoo S. , Kao T. , Dehqanzada Z. A. , Storrer C. E. , Harris K. A. , Jama Y. H. , Shriver C. D. , Ponniah S. , Peoples G. E.
Published: March 2020

10078

Background: Using the Luminex multiplex assay, we have reported significant correlations between serum levels of MCP-1, eotaxin, and IL-6 and disease characteristics in breast cancer (BCa) patients. We now examine the potential of utilizing a general cytokine profile to develop a statistical model to predict certain disease states in these patients. Methods: Sera from 36 BCa patients (24 node-negative, 12 node-positive, 12 normals) were analyzed using the Luminex assay for levels of 21 cytokines (IL-1α, 1b, 2, 4, 5, 6, 8, 10, 12, 13, 15, 17, IFN-γ, G-CSF, GM-CSF, TNF-α, IP-10, MIP-1α, RANTES, MCP-1, eotaxin). Logistic regression models were used to assess if a binary outcome variable (Y) can be predicted by using serum cytokine levels (X). The area under a receiver operating characteristic (ROC) curve (c) was used to assess the potential utility of a biomarker. The larger the value of c, the better the biomarker. Results: MCP-1 was found to be a possible predictor of the presence of BCa while other potential biomarkers were IL-13, MIP-1α and eotaxin. The higher the MCP-1 level, the greater the likelihood that the patient would have BCa. Similar relationships applied to the other potential biomarkers. Among BCa patients, GM-CSF seemed to be a good predictor of nodal status with lower levels of GM-CSF predicting positive nodes. Other potential biomarkers with a similar expression pattern for nodal status were MCP-1, IL-6 and IL-5. Due to small sample sizes, we were unable to examine a potential “panel” of cytokines to develop a prognostic algorithm based on serum analysis. Conclusions: MCP-1, which was previously shown to be elevated in BCa, may also have some predictive value linking the presence of disease and disease severity as measured by nodal status. Other prominent cytokines from earlier studies (MIP-1α, eotaxin, IL-6) also displayed some possible predictive value. Our results warrant studying a larger population in order to establish a unified prognostic formula for BCa based on serum cytokine levels.

[Table: see text]

No significant financial relationships to disclose.
Oxygen saturation measured in vivo in bronchial tumors during bronchoscopy in patients with lung cancer using differential path length spectroscopy (DPS) correlates with survival, stage of the disease, and expression of HIF1a
Authors: Aerts J. G. , Bard M. P. , Amelink A. , Sterenborg D. , Bom F. , Van Meerbeeck J. , Hoogsteden H. , Lambrecht B.
Published: March 2020

10084

Background: Tumor hypoxia is known to correlate with poor prognosis in solid cancers. In bronchial tumors direct determination of oxygen saturation is not possible and therefore indirect measures like hypoxia related proteins as HIF1a have been studied. We studied the prognostic significance on patient survival and tumor stage of microvascular oxygen saturation (StO2) measured in vivo in bronchial tumors using optical spectroscopy and compared this to HIF1a expression in bronchial biopsies. Methods: DPS is a non-invasive real-time measurement of hypoxia related parameters such as microvascular blood oxygenation (StO2) which can be used during bronchoscopy (Bard Am J Respir Crit Care Med 2005). Endobronchial tumors (12 squamous cell carcinomas, 5 adenocarcinomas, 15 large cell undifferentiated and 6 small cell carcinomas), were measured in 38 patients. In a subgroup of patients HIF1a expression was related to StO2 using linear regression analysis. Results: StO2 of bronchial tumors was predictive for patient survival. Patients with an endobronchial tumor and a saturation lower than the median value of 52% had a significant shorter overall survival than patients with a better saturated tumor. The StO2 was also related to the stage of the disease. StO2 was significantly lower in case of advanced lung tumors (stage III & IV, StO2= 44.7 ± 24.7%) in comparison with localized lung tumors (stage I&II, StO2 = 73.3 ± 12.4) (p<0.05). StO2 correlated to HIF1a expression levels (r = −0.65). Conclusions: StO2 using DPS correlates with HIF1a expression, is related to tumor stage and is a prognositc factor. This opens a new field for study of therapeutic agents in lung cancer and may also be important in the staging of lung cancer.

No significant financial relationships to disclose.
A multi-gene algorithm as predictor of response to chemo-radiotherapy in head and neck cancer
Authors: Gasco M. , Syed N. , Smith P. , Numico G. , Colantonio I. , Garrone O. , Granetto C. , Di Costanzo G. , Merlano M. , Crook T.
Published: March 2020

10086

Background: Chemo-radiotherapy results in clinical cure for stage III/IV head and neck cancer in approximately 40% of cases, with significant treatment-associated morbidity and mortality. Molecular genetic factors predictive of treatment outcome would clearly be of value in selection of patients with highest probability of response. We have analysed the structure and epigenetic regulation of specific genes as possible predictors of outcome to chemo-radiotherapy. The genes analyzed were: p53 (single nucleotide polymorphism (SNP) and mutation), MDM2 (SNP), Chfr (methylation), CRABP1 (methylation). Methods: 84 patients with locally advanced head and neck cancer receiving cisplatin-based chemo-radiotherapy were studied. SNP genotypes were determined by direct sequencing of DNA from normal tissue. Acquired mutations in p53 and aberrant methylation in the CpG islands of specific genes were analyzed by direct sequencing and methylation-specific PCR. Results: There were 6 treatment-related deaths in 84 patients, all occurring in cases with germ-line haplotype p53 72 Arg/Arg, MDM2 309T/T. At the time of analysis, 39/84 (46%) patients had progressed or died, with median time to progression or death = 17.3 months. Complete response was more common in patients whose genetic/epigenetic profile was p53 72 Arg/Arg wild type, MDM2 309 G/G, Chfr methylated, CRABP1 methylated, compared to the profile p53 72 Arg/Arg mutant, MDM2 309 T/T, Chfr unmethylated, CRABP1 unmethylated (91% vs 46%, log rank p = 0.002). Progression-free survival was significantly longer for patients with the profile p53 72 Arg/Arg wild type, MDM2 309 G/G, Chfr methylated, CRABP1 methylated, compared to those with p53 72 Arg/Arg mutant, MDM2 309 T/T, Chfr unmethylated, CRABP1 unmethylated (% surviving progression-free at 2 years = 74% vs 36%, p = 0.001). Overall survival was significantly longer in patients with the profile p53 72 Arg/Arg wild type, MDM2 309 G/G, Chfr methylated, CRABP1 methylated compared to those with p53 72 Arg/Arg mutant, MDM2 309 T/T, Chfr unmethylated, CRABP1 unmethylated (% surviving at 2 years = 88% vs 40%, p = 0.0004). Conclusions: Genetic and epigenetic parameters influence the toxicity and clinical outcome of chemo-radiotherapy in head and neck cancer.

No significant financial relationships to disclose.
TP53 status is an important prognostic marker for patients with hepatic metastases from colorectal carcinoma
Authors: Mollevi D. G. , Serrano T. , Ginesta M. M. , Valls J. , Torras J. , Ramos E. , Germà J. , Moreno V. , Capella G. , Figueras J. , Villanueva A.
Published: March 2020

10092

Background: The aim of this study was to analyze the value of TP53 gene mutations as a prognostic marker to improve the selection of patients candidates to surgery in a consecutive series of liver metastases from colorectal cancer patients. Methods: 91 patients with liver metastases from colorectal carcinoma (CRC) were included. Mutational study of TP53 gene, exons 4 to 10, was assessed both in paraffin-embedded hepatic metastasis and normal liver parenchyma by SSCP (Single Strand Chain Polymorphism) followed by sequencing of abnormal electrophoretic mobility patterns. Immunostaining of P53 and P21 proteins were assessed in the same group of patients. Results: Forty-eight out 91 (50.05%) metastases showed mutation in TP53. Higher incidence of mutations was detected in exons 5–8, although exons 9 and 10 were mutated in 28.26% of metastases. Protein-truncating mutations (nonsense and frameshift) occur in 47.8% of metastasis harboring TP53 mutations. TP53 mutation was associated with poor prognosis in univariate (P=0.0062) and multivariate Cox proportional hazard model (P=0.012) analysis. Prognosis association was maintained in the group of patients undergoing radical resection, named R0 series (n=79; P=0.008). High prevalence of TP53 mutations happen in patients with >3 metastases (65.6%; P=0.034), primary tumors Duke’s C-D stages (57.4%; 63.0%; respectively; p=0.026) and patients with age <57 years at resection. Interestingly, patients with TP53 mutations in their metastases relapsed earlier after the resection of the primary tumor (P=0.026). Postoperative 5-FU-based chemotherapy showed a better survival outcome in patients with wild-type TP53 hepatic metastases (HR: 2.92; 95% CI: 1.32–6.46; P=0.006). Conclusions: TP53 mutational status seems to be an important prognostic factor in patients with hepatic metastases from CRC.

No significant financial relationships to disclose.
ErbB3, Cyclin D1 and Ki67 nuclear staining predicts biochemical recurrence in prostate cancer treated by radical prostatectomy
Authors: Koumakpayi I. H. , Gannon P. O. , Le Page C. , Alam-Fahmy M. , Madore J. , Mes-Masson A. , Saad F.
Published: March 2020

10096

Background: The nuclear accumulation of growth factor receptor was reported to be associated to increased cell proliferation. Cyclin D1 and Ki67 are nuclear markers of cell proliferation. Deregulation of Cyclin D1 and Ki67 expression play a role in tumorigenesis and metastasis. Recently, we observed that nuclear localization of ErbB3 was associated with prostate cancer progression. The objective of this study was to determine if the association of cell proliferation markers and nuclear localization of ErbB3 could predict biochemical recurrence (BCR) in patients with prostate cancer following radical prostatectomy. Methods: Using immunohistochemistry we analyzed a tissue microarray containing 386 cores from 64 formalin-fixed paraffin embedded specimens from prostate cancer patients who had undergone radical prostatectomy. No patient had received hormone therapy prior to surgery and prior to BCR. Antibodies against Cyclin D1, ErbB3 and Ki67 proteins were used. Results: Nuclear staining was 60%, 67% and 86% for Cyclin D1, ErbB3 and Ki67 respectively. In our cohort, 29 of 64 PCa patients (45%) had a BCR after a median 3 years of follow-up. Thirty seven (37) percent of patients had positive nuclear staining for all three markers. BCR free survival probability at 3 years was not significant for each marker individually, except for ErbB3 in positive surgical margin patients. When all three markers were combined for nuclear staining Kaplan-Meier analysis BCR free was 0.4 and 0.1 for positive and negative nuclear staining respectively (p=0.0068). Univariate COX regression analysis shows a 2.98 fold (95% CI: 1.29 - 6.86, p=0.01) higher rate of BCR in patients positive for these three markers. In addition, in a multivariate model, including pre-operative PSA (p=0.19), pathologic stage (p=0.29), Gleason grade (p=0.40) and specimens that had positive nuclear staining for the 3 markers were associated with a 3.97 fold higher rate of BCR (95% CI: 1.54 - 10.25, p=0.0068). Conclusion: These results suggest that the association of cell proliferation markers and nuclear localization of ErbB3 could be useful in predicting recurrence following radical prostatectomy and guide therapeutic decisions. Large scale trials are needed to confirm these results.

No significant financial relationships to disclose.