bluuebunny/crossref_metadata_2025 huggingface dataset:
DJ-1 as a predictor of pathologic complete remission of neoadjuvant chemotherapy with breast cancer patients.
Authors: Kawate Takahiko
Published: March 2020

10544

Background: DJ-1 is a multifunctional protein which is encoded by the causative gene of familial Parkinson disease (i.e., PARK7), and is associated with carcinogenesis. DJ-1 is also a candidate marker for the presence of breast cancer cells, in which it is secreted in nipple fluid and serum. We previously reported that a DJ-1 expression pattern that is low at the protein level and high at the mRNA level is associated with DJ-1 secretion. This secretory expression pattern correlated with a negative hormone receptor status and with unfavorable clinical outcome in breast cancer patients. However, the association of the DJ-1 secretion pattern with therapeutic effect has yet to be determined. Methods: A total of 299 patients received neoadjuvant chemotherapy and surgery from 2002 to 2010 at our institution. We performed immunohistochemistry and in-situ hybridization of DJ-1 in both the needle biopsy and operative specimens of 147 cases with a follow-up time of 3.1 years. The average degree of staining intensity was evaluated semi-quantitatively using an image analyzer. Univariate and multivariate analyses were used to evaluate the predictive value of the therapeutic effect of neoadjuvant chemotherapy. Results: A low expression of DJ-1 protein was detected in the needle biopsy and operative specimens of 89 of the 147 cases, regardless of a high or maintained mRNA level. The tumor response was evaluated as pathological complete remission (pCR) in 39 cases and as non-pCR in the remaining 108 cases. A low expression of DJ-1 protein, which was detected in 34 (87.2%) pCR cases, was a significant predictor on univariate analysis (P <0.001). On multivariate analysis, a low expression of DJ-1 was shown to be a significant independent predictor similar to established predictors such as estrogen receptor status and human epidermal growth factor receptor 2 score. Conclusions: A low expression of DJ-1 protein can serve as a novel and important predictor of a neoadjuvant chemotherapeutic effect, as well as a promising indicator for serologic diagnosis in breast cancer patients.
Evaluation of CYP3A5, VEGF-a, and VEGFR2 polymorphisms as markers of sunitinib toxicity.
Authors: Rodriguez de Antona Cristina , Grande Pulido Enrique , Leandro-Garcia Lj , Barriuso Jorge , Capdevila Jaume , Jimenez Paula , Castelo Beatriz , Castellano Daniel E. , Longo Federico , Garcia-Donas Jesus
Published: March 2020

10546

Background: Sunitinib (Sun) is a multitargeted tyrosine-kinase inhibitor approved for the treatment of renal cell carcinoma (RCC), GIST and pancreatic neuroendocrine tumors (pNETs). It is known that a polymorphism in Sun metabolizing gene CYP3A5, was associated with an increased risk of dose reductions due to toxicity (tox). Additionally, polymorphisms in VEGF-A and VEGFR2 were suggested to increase the risk of hypertension (HT) during treatment (García-Donas, et al. Lancet Oncol 2011). These data have not been validated so far. We aim to assess the value of these polymorphisms as markers of tox using a wide range of solid tumors treated with Sun. Methods: In this non-interventional and retrospective study, DNA was collected from 28 patients with different pathologies (16 pNETs, 5 medullary thyroid carcinomas, 2 NETs of the lung, 2 undifferentiated follicular carcinomas, 1 undifferentiated papillar carcinoma, 1 GIST, and 1 carcinoid of the rectum) treated with Sun in a daily practice setting. Genotyping for CYP3A5*1 allele (rs776746), VEGF-A -2578C>A (rs699947) and VEGFR2 Q472H (rs1870377) was performed. Associations between the genotypes and Sun dose reductions and HT were performed using univariable analyses. Results: In agreement with previous reports, we found that VEGF-A rs699947 conferred a statistically significant increased risk of developing HT during treatment (HR=9.8, 95%CI=1.2-82.0, P=0.034). CYP3A5*1 high metabolizing allele showed a trend towards the increase in the risk of Sun dose reductions due to tox (HR=2.4, 95%CI=0.8-6.8, P=0.11) with a median time to dose reduction of 7.0 months for wild type homozgygous patients and 4.6 months for heterozygous patients. A trend was also found on the relation between VEGF-A rs699947 A/A patients (HR=3.8, 95%CI=0.8-16.8, P=0.080) and dose reduction risk. No significant associations were found for VEGFR2 rs1870377. Conclusions: The present study suggests that VEGF-A rs699947 is a risk factor for Sun HT and has a role in other tox leading to dose reductions. Similarly, CYP3A5*1 shows a trend towards an increased risk of Sun dose reductions. If confirmed, these markers could be used to identify a subset of patients with an increased risk of Sun toxicity.
Pharmacogenetic predictors of adverse events in stage II-III colon cancer (CC) patients treated with oxaliplatin and fluoropyrimidines-based adjuvant chemotherapy (CT): A GEMCAD study.
Authors: Custodio Ana , Moreno Juan , Aparicio Jorge , Gallego Plazas Javier , Fernandez-Martos Carlos , Yaya Ricardo , Calatrava Ana M , Maurel Juan , Burgos Emilio , Tejerina Eva , Barriuso Jorge , Moreno Victor , Martinez Marín Virginia , De Castro Javier , Lamarca Angela , Lapunzina Pablo , Madero Rosario , Cejas Paloma , Feliu Jaime ,   Grupo Español Multidisciplinar de Cáncer Digestivo (GEMCAD)
Published: March 2020

10547

Background: Although the benefit of oxaliplatin-based adjuvant CT has been demonstrated in patients with resected stage II-III CC, the recommendation to administer postoperative treatment must consider its potential adverse events. Predicting individual patients’risk of severe toxicity could potentially improve the quality of care by allowing individualization of treatment. We investigate the utility of determining single nucleotide polymorphisms (SNPs) in genes involved in oxaliplatin and fluoropyrimidines metabolisms to predict the toxicity of adjuvant CT in stage II-III CC patients. Methods: DNA was extracted from formalin-fixed paraffin-embedded samples from 379 surgically treated high-risk stage II (27.71%) and stage III (72.29%) CC patients receiving adjuvant CT (54.35% FOLFOX, 45.65% XELOX) from January 2004 to December 2008. Genotyping was performed for 35 SNP in 18 genes using the MassARRAY (SEQUENOM) technology. Results: A total of 89 (23.4%) patients experienced at least one grade 3-4 adverse event. The most common grade 3-4 toxicities were neutropenia (15.3%), diarrhea (8.17%) and neurotoxicity (7.65%). The MTHFR rs1801133 C>T C/C (30.1% C/C, 17.5% C/T, 21.7% T/T; p=0.043) and XRCC2 rs3218408 T>G T/T (28.3% T/T, 16.2% G/T and 10% G/G; p=0.007) genotypes were associated with higher risk of any grade 3-4 toxicities, whereas the incidence of severe toxicity was lower in patients with the UMPS rs3772807 G>C C/C (14% C/C, 21% C/G, 28.9% G/G; p=0.021) and DPYD rs970337 G>A A/A (16.7% A/A, 21.1% A/G, 30.1% G/G; p=0.028) genotypes. In addition, the DPYD rs970337 G>A A/A genotype was associated with a lower rate of grade 3-4 diarrhea (5% A/A, 9.4% A/G, 8.1% G/G; p=0.030), the ABCG2 rs3114018 A>C C/C genotype with an increased rate of grade 3-4 neutropenia (15.2% C/C, 6.3% A/C, 4.5% A/A; p=0.034) and the ERCC1 rs11615 T>C T/T genotype with a lower rate of grade 3-4 neurotoxicity (5.4% T/T, 9.1% C/T, 10.2% C/C; p=0.032). Conclusions: Our data suggest that SNPs in genes involved in oxaliplatin and fluoropyrimidines metabolisms can potentially predict severe toxicity in stage II-III CC patients treated with adjuvant CT.
Use of gene expression and alternative splicing signatures to discriminate breast cancer stem cells from fibroblasts.
Authors: Weaver David T. , Shapiro Irina M , Derr Alan G , Paterson Daniel , Pachter Jonathan A
Published: March 2020

1057

Background: Tumors frequently contain cancer stem cells (CSCs) or tumor-initiating subpopulations, with an ability to self-renew and regenerate all cell types within the tumor. Basal-like breast cancers exhibit features of CSCs, including expression of surface markers, even though these cells are rare. Given the role of CSCs in the recurrence and spread of cancer, there is an urgent need to develop new therapeutic agents that target CSCs. Development of CSC-targeted drugs will be greatly facilitated by biomarkers that can identify CSCs to aid in patient selection and determination of drug response. Defining the CSCs in tumors is complicated by the high mesenchymal nature of fibroblasts. Analysis of gene expression and alternative splicing patterns in CSCs that are not observed in fibroblasts may provide valuable new CSC-specific markers. Methods: Alternative splicing and gene expression microarray strategies were used to identify selected exons and differentially expressed genes between 10 Basal human breast cancer cell lines and a combination of 12 Luminal and 3 fibroblast cell lines. Q-PCR analysis was conducted to determine candidate CSC gene differential expression between Basal, Luminal, and Fibroblast cells lines. Results: Expression levels of 11 genes were higher and 24 genes were lower in the Basal cell lines versus Luminal or fibroblastic cell lines. Comparison of Basal cell lines to the Luminal/Fibroblast cell lines identified 36 cassette exons that were included, and 26 that were excluded in Basal cell lines. Also, 19 genes were upregulated in Basal cell lines compared to the other groups as detected by Q-PCR. Interestingly, the 19-multigene model defined the Triple Negative Breast Cancer patients that were Likely to Recur under standard chemotherapy with a p = 1.90e-03 and AUC 0.723. Conclusions: Gene and exon marker sets distinguish CSC versus fibroblasts and may be instructive in identifying patients that recur early in Triple Negative Breast Cancer. The CSC-associated RNA signatures identified here will be further refined to develop new CSC-specific diagnostic markers to stratify breast cancer patients and monitor response to novel CSC-targeted therapies.
FDG-PET and CT as early predictors of outcome in NSCLC patients treated with erlotinib.
Authors: Iannone Robert , Evelhoch Jeffrey L. , Lam Raymond , Bergstrom Donald Alan , Thornton Bob , Herman Gary A. , Mozley P. David
Published: March 2020

10573

Background: Tumor metabolism as assessed by FDG-PET is an early biomarker for clinical benefit in oncology. This multi-national study evaluated associations of changes in mean standardized uptake values (SUVmean) of FDG and tumor burden with disease control rates (DCR: CR+PR+SD), PFS and OS in 48 NSCLC patients (EGFR mutants or favorable clinical/demographic characteristics) treated with erlotinib. The primary hypothesis was that changes in SUVmean at day 8 predicted DCR. Methods: Patients were scanned by FDG-PET (twice pre-therapy, 1 and 3 weeks post-therapy) and CT (same as FDG-PET and every 6 weeks post-therapy until progression or death). Early response biomarkers included the decrease in SUVmean (DiSUV), sum of diameters (DiSOD) and sum of volumes (DiSOV). For the primary hypothesis, metabolic response (MR) was defined as a decrease of >2SD the variability between baseline PET scans (30%). Exploratory analyses of the biomarkers at various thresholds were performed. Results: The primary hypothesis was met: DCR was 34.6% (80% CI: 17.5%, 48.2%) higher in patients with a MR at day 8 (p = 0.033); the association was stronger for day 22. The table compares hazard ratios derived from the "optimal" % decrease (smallest upper 80% confidence limit) over the 10% to 50% (by 10%) range for all biomarkers and time points, by PFS and OS. Conclusions: FDG-PET was predictive for clinical responses after 8 days of treatment. Comparing all biomarkers, DiSOV had stronger associations than both DiSUV and DiSOD with PFS or OS. For NSCLC treated with erlotinib in this clinical context, the decrease in CT-based tumor volume may be a better early predictor of clinical outcome than FDG-based SUVmean. [Table: see text]
Preoperative detection of circulating tumor cells in patients with colorectal carcinoma: Correlation with clinicopathologic variables and recurrence.
Authors: Sakihama Hideyasu , Kobayashi Nozomi , Funakoshi Tohru , Shimokuni Tatsushi , Homma Shigenori , Takahashi Norihiko , Kataoka Akihiko , Ozaki Michitaka , Kamiyama Toshiya , Taketomi Akinobu
Published: March 2020

10578

Background: The role of circulating tumor cells (CTC) in the management of colorectal cancer patients has not fully established. The aims of this study are to investigate the relationship between the presence of CTC with clinicopathological variables and recurrence by magnetic activated cell sorting (MACS) system. Methods: Peripheral blood samples were collected from 80 patients. Enrichment of CTC was performed by direct immunomagnetic labeling of EpCAM positive cells in peripheral blood. Subsequently, double immunofluorescence for cytokeratin and CD45 was performed to detect CTC. Peripheral blood samples from twenty healthy volunteers were used as controls. Results: Preoperative positive rate of CTC was 35% while specificity was 100%. No CTC was found in peripheral blood from healthy volunteers. No correlation was found between the presence of CTC and location of tumors, grade of differentiation, vessel invasion, lymph node metastasis or TNM stages. On the other hand, the depth of invasion (0% in Tis, 11.1% in T1, 18.2% in T2 and 34.7% in T3+T4, P=0.05) and tumor recurrence (28.2% for initial operation and 88.9% for reoperative surgery for tumor recurrence, P<0.001) closely correlated with the presence of CTC. Preoperative positive rate of CTC among patients who have recurred postoperatively was 75%. Conclusions: Our results indicate that detections of CTCs correlate with the depth of invasion and tumor recurrence. Preoperative presence of CTCs might be a strong predictor for tumor recurrence.
Antigens diagnoses in paraneoplastic neurological syndromes.
Authors: Rudnicka Halina , Jagiello Gruszfeld Agnieszka I. , Glogowska Iwona
Published: March 2020

10582

Background: At present, paraneoplasmatic neurological syndroms (PNS) are thought to be autoimmunological disorders. Neurological symptoms in PNS result from the disturbance of the nervous system, with a participation of the immune response triggered by the aberrant expression of the so-called onconeuronal antigens of a tumour. Very important for PNS diagnostics is the investigation of antineuronal antibodies (ANA) in the serum. Different types of ANA are frequently associated with specific tumors and with specific neurological syndromes.Investigations of serum ANA in patients with breast cancer with the symptoms of PNS. Methods: Investigations of ANA in serum were performed with the indirect immunofluorescence test but the type of ANA was analyzed by the Western Blot (WB) method . The presence of these antibodies which are included to the group of “the so called well characterized” is believed to be a definite (95 – 100%) determinant of the PNS diagnosis. Results: We examined 147 patients with breast cancer suspected to have PNS, hospitalized at Breast Cancer Clinic of the Institute of Oncology in Warsaw. The immunofluorescence test was positive in 92 patients, however in 24 patients the WB occurrence of “the well characterized” antibodies was found. In this group we observed: in 4 patients the cerebellar degeneration (anti-Yo, anti-Ma2); in 6 patients brainstem encephalitis and encephalomyelitis combined with sensory neuropathy (anti-Ma2, anti-CV2, anti-Yo, anti-Hu, anti-Ri); 4 myastenic syndrome (anti-Hu,anti-Ri,anti-Ma2/Ta); 5 sensory neuropathy (anti-Hu, anti-Yo, anti-Ma2, anti-Ri); and in single cases we also observed polyneuropathy: Guillain-Barre (Ma2): Miller-Fisher syndrom (anti-Ma2/Ta); cancer-associated retinopathy (anti-Yo); dermatomyositis (anti-Ri) and scleroderma (anti-Yo). Conclusions: Our analysis indicates that the investigation of the presence of ANA in the serum is the important in the diagnosis of PNS in patients with breast cancer. The types of ANA correspond to the individual paraneoplastic neurological syndromes which opens the proper methods of treatment in PNS.
Estimation of expected survival time using gene expression profiling for tumor site origin.
Authors: Henner William David , Nystrom J. Scott , Hainsworth John D. , Varadhachary Gauri R. , Greco Frank A. , Amin Mahul , Walker Michael , Becker Shawn , Pillai Raji , Stein Sandy , Hornberger John C.
Published: March 2020

10584

Background: Expected survival is an important factor in treatment selection and patient decision making. Gene expression profiling for tissue of origin (GEP TOO) can be used to classify metastatic and poorly-differentiated tumors according to the most likely primary site for difficult-to-diagnose tumors. The Pathwork Tissue of Origin Test, (Redwood City, CA) generates Similarity Scores (SS), one for each of the 15 tissues on the Tissue of Origin Test Panel and sum to 100. The highest SS has been validated as the most likely tissue of origin in a large validation study (Pillai et al. 2010). This abstract reports the relationship between the highest SS for a sample and patient survival. Methods: Two cohorts were analyzed in IRB-approved studies. Cohort 1 consisted of 45 patients with carcinoma of unknown primary treated empirically with therapy for carcinoma of unknown primary with known outcome data including survival. Cohort 2 consisted of 107 patients receiving the TOO test in routine clinical practice and included in a decision impact registry study. The relationship between the highest SS generated by the TOO test was examined using parametric (Gamma) and non-parametric (Cox Proportional Hazards) regression models with covariates of age, gender and median survival of the cancer type indicated. Logrank tests were performed to compare survival in patients with SS above or below 50 in each cohort. Results: The highest SS generated by the TOO Test was strongly and positively correlated with patient survival in both Cohort 1 (p = 0.0093, Cox PH) and in Cohort 2 (p=0.0045, Gamma). In Cohort 1, survival for patients with a SS < 50 was 5.9 mos and for patients with a SS ≥ 50 was 9.7 mos (p=0.03, logrank). In Cohort 2, survival for patients with a SS < 50 was 10.6 mos and 22.6 mos for patients with a SS ≥ 50 (p=0.0073, logrank). Conclusions: The Similarity Scores generated by GEP TOO Test have been validated as an aid to diagnosis for difficult-to-diagnose malignancies. Two independent studies of this GEP TOO Test demonstrated a strong positive relationship between the highest SS and survival. This has the potential to expand the clinical utility of this test beyond diagnosis to include prognosis. Further studies to examine this question are underway.
Novel malignant-mesothelioma-associated antigens (Gene-X and THBS-2) in the diagnosis of malignant pleural mesothelioma (MPM).
Authors: Tanaka Fumihiro , Shigematsu Yoshiki , Hanagiri Takeshi , Uramoto Hidetaka , So Tomoko , Yoneda Kazue , Takuwa Teruhisa , Hashimoto Masaki , Matsumoto Seiji , Okumura Yoshitomo , Kondo Nobuyuki , Hasegawa Seiki
Published: March 2020

10585

Background: Malignant pleural mesothelioma (MPM) is a highly aggressive malignant tumor of the pleura associated with asbestos exposure, and its diagnosis is usually difficult at early stage. We identified novel mesothelioma-related antigens, Gene-X and thrombospondin-2 (THBS-2), recognized by tumor-infiltrating B cells (Cancer Sci 2009), but the clinical significance in the diagnosis of MPM remains unclear. Methods: A total of 120 patients, who presented with a suspicion of MPM and received pleural biopsy, were reviewed; 97 patients were finally diagnosed with MPM and 27 were with non-malignant diseases (NM). The antibody-titers against Gene-X and THBS-2 in the sera were measured by ELISA method. Results: The serum antibody-titer against THBS-2 was significantly higher in MPM patients than in NM (P<0.01), but there was no difference in the serum antibody-titer against Gene-X (Table). The receiver operating characteristic (ROC) curve analysis showed a significant diagnostic value of serum antibody-titer against THBS-2 with the area-under curve of 0.886 (95% CI, 0.797 - 0.975; P<0.001) in discrimination of MPM from NM diseases; the sensitivity and specificity, when the cut-off value was 0.08, were 72.2% and 95.5%, respectively. Conclusions: The serum antibody-titer against THBS-2 can be a useful non-invasive marker in the diagnosis of MPM. [Table: see text]
Multisite validation of a 92-gene molecular classifier: Diagnostic accuracy and clinical utility in metastatic cancer.
Authors: Schnabel Catherine A. , Kerr Sarah E , Sullivan Peggy S , Zhang Yi , Singh Veena , Carey Brittany , Erlander Mark G. , Highsmith W Edward , Dry Sarah M , Brachtel Elena F
Published: March 2020

10588

Background: Accurate tumor classification is increasingly critical to individualized care as patient outcomes improve with use of targeted cancer therapies and predictive biomarkers. In metastatic cancers of uncertain or unknown origin, identification of a primary site remains equivocal in a significant number of cases. Gene expression signatures may improve accuracy and specificity of tumor classification, however, large-scale, blinded validation studies that demonstrate stable performance in metastatic and poorly differentiated cases are currently lacking. Methods: Seven hundred and ninety cases (51% female, 44% metastatic, 63% grade 2 and 3, 14% limited tissue specimens) representing 28 tumor types and 50 subtypes were processed and adjudicated between Mayo Clinic, UCLA and Massachusetts General Hospital. Blinded FFPE tumor sections were submitted and tested using a 92-gene RT-PCR assay (CancerTYPE ID, bioTheranostics Inc.). Molecular predictions were evaluated for concordance with the reference diagnosis; diagnostic accuracy between clinical subsets was compared. Results: The 92-gene assay demonstrated overall sensitivities of 87% for tumor classification and 82% for subtyping. Forty-seven (5.9%) cases were considered unclassifiable. Comparative analysis of performance between metastatic (n=329) and primary (n=414) tumors showed no statistically significant difference in accuracy (85% vs 88%, p=0.16). Similarly, no significant decreases in performance were observed across histological grades (p=0.58) and when comparing limited tissue to excisional biopsy specimens (p=0.16). Strong precision was demonstrated for accurate identification of a primary tumor in tissues biopsied from common metastatic sites, reported as positive predictive values of 100% for lung, brain and peritoneum, 92% for ovary and 80% for liver. Conclusions: Results from this multisite, blinded study validate the diagnostic accuracy of the 92-gene assay for classifying a diverse set of tumors, and support its clinical utility in the diagnosis of metastatic tumors to determine tissue of origin, and in the differential diagnosis of metastatic vs new primary disease.
Genomic profiling of non-small cell lung cancer (NSCLC) for personalized targeted therapy using CT-guided transthoracic needle biopsy (TTNB).
Authors: Gill Ritu R. , Heon Stephanie , Yeap Beow Yong , Butaney Mohit , Lindeman Neal Ian , Rabin Michael S. , Jackman David Michael , Janne Pasi Antero , Johnson Bruce E. ,   Dana-Farber Cancer Center
Published: March 2020

10592

Background: Genomic profiling for personalized targeted therapy is emerging for NSCLC. DFCI introduced systematic testing for mutations in BRAF, HER2, PIK3CA and ALK translocations in addition to EGFR and KRAS in July 2009 as part of a prospective study. We report the utility, efficacy and safety of CT guided TTNB in this cohort. Methods: Patients with stage IV or relapsed NSCLC seen at the DFCI were referred to BWH for CT guided TTNB of their tumors to identify driver mutations prior to starting therapy. Pathology specimens were dissected and analyzed by PCR-Sanger sequencing for mutations in selected exons of EGFR, KRAS, BRAF, PIK3CA and HER2. ALK rearrangements were detected with fluorescence in-situ hybridization (FISH). Testing was performed after the pathologist deemed that the tissue was adequate. Complications such as pneumothorax and hemorrhage were recorded. Admission rates were also recorded. Results: Between 7/1/2009 and 1/09/2011, 81 patients underwent TTNB. The median age was 63 years. 54 (67%) were female, 66 (88%) were former/current smokers and 58(72%) had stage IIIB/IV disease. 64(79%) patients had sufficient tissue on core biopsies for genomic profiling, 4 (6%) of the 64 patients failed analysis for ALK rearrangements due to less than 50 tumor cells on the hybridized slide. The number of samples obtained ranged from 1-5 (2 cm 18-20 (G)). Lesions biopsied ranged in size from 1.2–8.9 cm. Mutations were identified in 38/81 (46.9%) patients (EGFR: 18; KRAS: 17; ALK: 2; PIK3CA: 1). 23(28.3%) had pneumothoraces 15(<10%), 5 (10-30%) and 3(>30%). 6 (7%) patients needed chest tubes. 9 (11%) were admitted post procedure for observation (8 for (24hrs) and 1 (72hrs). 19(23%) (18 grade1; 1 grade 2) had intra-parenchymal hemorrhage. A higher rate of pneumothorax was observed with the 18 gauge needles (p =.05). 15 of 20 (75%) patients with EGFR, HER2, BRAF or ALK alterations were treated with molecularly targeted therapy based on their genetic alteration. Conclusions: CT guided TTNB is a feasible, safe and efficacious technique for genomic profiling for targeted therapy and enables the acquisition of sufficient tissue for gene mutation analyses.
A circulating epithelial cell type merging stem cell, EMT, and hypoxic stress markers in early and advanced breast cancer.
Authors: Habets Leo , Koerber Wolfgang , Frenken Bettina , El Ghali Ilham , Danaei Mahmoud , Kusche Manfred , Peisker Uwe , Kroll Torsten , Pachmann Katharina
Published: March 2020

10600

Background: MAINTRAC as developed by our coworkers from Jena finds more CETC. They found CETC in 90% of high risk patients. Circulating cells in epithelial mesenchymal transition (EMT+) should be detectable in early disease. Methods: 2 colours (7AAD, EP-FITC) detected living and dead cells. Cells in EMT (EP-FITC, Vim-PE) were analysed in paralell. Complex marker expression urged us to introduce a three colour technique. 1. EPCAM-FITC, Vimentin-PE and DAPI , marking living or dead cells in EMT. 2. EP-FITC, Vim-PE and CD44 PB for stemness. ACA9 (hypoxia) and PARP-1 expression (genotox) were used too. Results: In the 2 colour phase we examined CEC in 110 patients with early disease and 44 patients with metastasis. Cells were detectable in both disease situations in 50 and 60% of patients, in a control population only in few patients low numbers were found. In non cancer situations e.g liver disease high numbers of EMT+cells were found.The results of 3 colour analysis are shown in the table. No clear differences between the classical risk groups are found, other subgroups are too small yet to be considered. Conclusions: Our findings show that in both disease situations cells merging EMT, stemcells ( EP+,Vim+.CD44+) and other markers are detectable. This cell type however is found also in non cancer conditions.We hypothesize that hypoxia is the common driver. Metastasis gives different patterns. Highly aggressive cancers show often none or few cells. In less agressive disease high cell numbers are found. We need to characterize the "benign" EMT phenomenon, as it could mingle with "real" CTC. Definitely liver is the source of these benign cells. Multi colour analysis will reveal the differences. Nevertheless monitoring of this special cell type could give new insights in the metastatic process in early and late disease. [Table: see text]
Exploratory study of histopathological and biomolecular characteristics of breast cancer in two different populations: African (Tanzania) and Caucasian (Italy).
Authors: Serra Patrizia , Bravaccini Sara , Masalu Nestory Andrew , Puccetti Maurizio , Rengucci Claudia , Tumedei Maria Maddalena , Serra Luigi , Zoli Wainer , Amadori Dino ,   on behalf of Associazione Vittorio Tison
Published: March 2020

10601

Background: There is little information available on the clinical and molecular epidemiology of breast cancer in Sub-Saharan African women. In Tanzania, breast cancer mortality is the second cause of death after cancer of the uterine cervix, with the majority of tumors diagnosed at a very advanced stage. We aimed to compare the distribution of histopathological and biomolecular characteristics (ER, PgR, Ki67, HER2, grading) of breast cancer in African (Tanzania) and Caucasian (Italy) patients. Methods: 142 women with invasive breast cancer, 71 from Tanzania and 71 from Italy, were considered. Histopathological classification and biomolecular determinations were performed at the Biosciences Laboratory of the Cancer Institute of Romagna (I.R.S.T., Meldola, Italy) as part of a joint Cancer Control Project between I.R.S.T and the Bugando Medical Center in Mwanza, Tanzania. All cases were evaluable for histopatological characterization, while biomolecular determination was possible in only 61 of the African patients. Hormone receptor status and Ki67 were determined by IHC; HER2 was evaluated by IHC for the African population and by FISH for the Italian one. Results: Whilst histopathological patterns were similar in the two populations, some biomolecular characteristics differed substantially. Of note, 64% of African breast cancer patients had ER negative tumors compared to only 17% of the Caucasian population. Important differences were observed in high Ki67 (>15%) values: 76% in Tanzanian women vs 58% in Italian breast cancer cases. HER2 positivity rates also differed in the two populations (25% in Tanzanian and 17% in Italian patients), as did the incidence of triple negative disease (13% and 7% in African and Caucasian populations, respectively). Conclusions: This preliminary study shows that breast cancer in African (Tanzanian) women is characterized by a high frequency of HER2 positive, ER negative, highly proliferating tumors. These biological patterns, together with very advanced stage at diagnosis, could be considered the main prognostic factors related to the high mortality rates for breast cancer in this African population.
pAKT expression in paraffin-embedded xenograft tumors after fixation delays and human breast cancer by optimized immunohistochemistry.
Authors: Yang Sherry X. , Nguyen Diana , Rubinstein Larry , Sherman Mark E , Swain Sandra M. , Tomaszewski Joseph E. , Doroshow James H.
Published: March 2020

10603

Background: With progress in targeting PI3K/AKT pathway in the treatment of a wide range of cancers, it is getting more important than ever for optimization of detection method of AKT kinase, a central effector of the pathway, in archived tissues. Recently, we found that pAKT-S473 (pAKT) significantly predicts paclitaxel benefit in breast cancer (JCO 28: 2974, 2010), in which a good analytical and clinical performance of quantitative pAKT immunohistochemistry (IHC) was demonstrated. The current study evaluates pAKT stability in human breast cancer xenograft tumors after delays in fixation, and assesses a detection window following fixation delays with an established cutoff [staining index (SI) > 2]. Methods: Xenograft tumors with high (MDA-MB-468) and intermediate levels (MDA-MB-231) of pAKT were fixed or snapped frozen in 10% neutral-buffered formalin or liquid nitrogen after delays post-excision. pAKT staining was performed in xenograft tumors and 96 cases of human surgical breast tumors by our optimized and traditional IHCs, and Western blot. Results: The mean SIs were 133, 121, 112, 94, 84, and 66 using optimized, and in contrast were 51, 44, 29, 14, 12, and 6.4 on traditional method at 0, 15, 30, 60, 120 and 180 min (2.6-fold by comparing the optimized with the traditional at baseline) in MDA-MB-468 xenograft tumors. pAKT level was ½ by the optimized, similar to the level detected by Western blot, relative to 1/8 of the baseline by the traditional (10.6-fold) at 180 min. The logarithmic decline rate by the optimal was 3.1 times (95% CI, 2.4 - 3.7) less than that of the traditional (normal approximation; 2-sided P < 0.0001). pAKT expression was observed in 38.5% (37/96 ) of surgical breast tumors, comparable to 38% (606/1581) in a large cohort derived from the NSABP B-28 trial. There was little loss of pAKT in MDA-MB-231 xenograft tumors up to 180 min by IHC and Western blot. Conclusions: The optimized pAKT IHC significantly increases the sensitivity of detection with a large window for positivity. It is suitable for use in archived human specimens although it warrants further standardization and validation among research laboratories and perhaps diagnostic laboratories in the future.
Monitoring of plasma pro-GRP level during EGFR-TKI treatment.
Authors: Kawano Yuko , Horiike Atsushi , Tanimoto Azusa , Sakatani Toshio , Saito Ryota , Kaburaki Kyohei , Yanagitani Noriko , Kudo Keita , Ohyanagi Fumiyoshi , Hagiwara Sachiko , Horai Takeshi , Nishio Makoto
Published: March 2020

10604

Background: Lung cancers harboring mutations in the epidermal growth factor receptor gene (EGFR) respond to EGFR tyrosine kinase inhibitors (EGFR-TKI), but drug resistance invariably emerges. The major acquired mechanisms of resistance are the EGFR T790M mutation or MET gene amplification. Transformation from NSCLC into small-cell lung cancer (SCLC) has been recently identified in acquired resistance to EGFR-TKI. However, it is difficult to predict the transformation during EGFR-TKI treatment because obtaining serial and sufficient specimens for biopsy is difficult. Pro-gastrin-releasing peptide (Pro-GRP) is a specific and sensitive tumor marker for SCLC. We evaluated the plasma Pro-GRP levels in EGFR-mutant NSCLCs and determined whether plasma Pro-GRP levels could predict SCLC transformation in resistance to EGFR-TKI. Methods: From July 2008 to December 2011, 49 patients with EGFR-mutant NSCLC who received EGFR-TKI treatment were enrolled. Plasma was obtained from these patients before EGFR-TKI treatment and when EGFR-TKI treatment failed. Pro-GRP and CEA levels were measured and compared before and after treatment. Results: Patient characteristics for 49 patients (15 men, 34 women) were as follows: median age, 62 years (41–81 years); histology, 46 adenocarcinomas (AD) and 3 non-AD tumors; and EGFR mutation type, 25 exon 19 deletions and 24 exon 21 L858R. All 49 patients had received EGFR-TKI treatment (45 with gefitinib and 4 with erlotinib); the response to EGFR-TKI treatment was PR in 39 patients, SD in 7, PD in 2, and NE in 1. Positive rate of ProGRP and CEA at pre-EGFR-TKI treatment was 2.0% and 57.2% and that at post-EGFR-TKI treatment was 6.1% and 69.4%, respectively. In 3 of 49 patients, the Pro-GRP levels had increased after treatment, but the CEA level did not increase. Objective responses to cytotoxic chemotherapy were noted in all 3 patients after EGFR-TKI treatment. Conclusions: Monitoring of plasma Pro-GRP during EGFR-TKI treatment may be useful for early detection of SCLC transformation in resistance to EGFR-TKI.
Study of the usefulness of angiotensin type 1 receptor (AGTR1) as a possible response predictor in patients with metastatic breast cancer (mBC) subjected to chemotherapy and treatment with bevacizumab (AVALUZ Study).
Authors: De la Haba- Rodriguez Juan , Salvador Bofill Javier , Codes Manuel , Ciruelos Eva M. , Jaen Ana , Galan Antonio , Gil Miquel , Serrano M. Jose , Baena Jose Manuel , Villanueva Silva Maria Jose , Gonzalez Encarnacion , Murias Adolfo , Miguel Ana , Garcia Helena , Bayo Juan L. , Sánchez-Rovira Pedro
Published: March 2020

10608

Background: Angiotensin type 1 receptor (AGTR1) is a membrane receptor implicated in the regulation of arterial pressure. It has also been related to carcinogenesis and tumor spread, and participates in neoangiogenesis. AGTR1 is expressed in a number of neoplasms as BC. In a previous neoadjuvant study, a significant relationship was observed between AGTR1 expression and CR to a regimen of chemo and bevacizumab (B). To study and confirm the relationship between tumor expression of AGTR1 and response to a regimen of chemo and B in pts with mBC. Methods: AGTR1 expression was subjected to immunofluorescence (monoclonal AGTR1 sc1173 antibody, Santa Cruz Biotechnology, CA, USA Dilution 1:50) in 50 samples of mBC pts from the AVALUZ study treated with first line bevacizumab 10 mg/kg, paclitaxel 150 mg/m2 and gemcitabine 2000 mg/m2 d 1 and 15 c/28 d until PD, unacceptable toxicity or medical decision. Immunoreactivity was reported as negative (0), low positive (1+) and high positive (2+). RNA was extracted from paraffin blocks, using the QIAamp DNA FFPE Tissue Kit and Deparaffinization Solution (QIAGEN) and expression of AGTR1 candidate gene was quantified using reverse transcription-PCR (RT-PCR). Results: 60% of the samples expressed AGTR1 (26% weak and 34% intense) – expression being more frequent in HR + tumors (65% vs 45%). Among the 50 samples analyzed, measurable disease was present in 39 pts. Treatment activity was significantly greater in the tumors AGTR1 expression 2+ (p=0.009) (Table). With a median follow-up of 19.30 months, the progression-free interval was longer in the tumors with intense AGTR1 expression (17.7 vs 10.8 months, p=0.132). More consolidated PFS and OS will be presented. Conclusions: AGTR1 expression may be useful as a predictor of response to chemo and bevacizumab. Prospective studies are needed to confirm these findings. [Table: see text]
The lipid metabolome of clear cell renal cell carcinoma (CCRCC).
Authors: George Saby , Choi Kyoung-Soo , Pili Roberto , Kazim Abdul Latif
Published: March 2020

10609

Background: The commonest type of kidney cancer is CCRCC. Treatment approaches mostly target aberrant vasculature. However, kidney cancer is also known to accumulate lipids and a detailed knowledge of the lipid species present in these tumors could lead to a better understanding of the underlying aberrant metabolic pathways and suggest possible treatment strategies. Lipidomics is an emerging field driven by rapid advances in mass spectrometry (MS), and is widely used to discover biomarkers. We attempt to identify the lipidomic profile of CCRCC using a liquid chromatography MS-based approach (LC-MS). Methods: We utilized 6 fresh frozen representative samples of CCRCC and matching non-tumor areas of kidney from nephrectomy samples. Lipids and other non-polar cellular constituents were extracted from both CCRCC and control tissues by methyl-t-butyl ether /methanol. LC-MS based lipid profiling was performed on a Waters Q-ToF Premier MS coupled with Ultra Performance LC. The peak detection and alignment across all chromatograms were performed using the XCMS software (v 1.14.1, Scripps Center for Metabolomics). Statistical comparisons of the intensities of aligned peaks were performed using the XCMS-built-in Welch's t-test. Results: The outcome of XCMS was converted to a table that contains fold change, p-value and mass to charge ratio (m/z) for each peak, its corresponding retention time, and the integrated peak intensities from all samples. 224 peaks out of 1419 differed between CCRCC and the control group, with p <0.05, calculated by XCMS. About an equal number of analytes increased or decreased in CCRCC compared with control samples. Preliminary attempts to identify the analytes included use of METLIN (Scripps Center for Metabolomics) and HMDB (Human metabolome database, Genome Alberta & Genome Canada) databases. Many of the hits identified phosphatidylcholines, phosphatidylethanolamines, triacylglycerols and diacylglycerols, as well as other lipid species. Conclusions: The lipid metabolomic profile varied significantly between CCRCC and control. Further studies are required to confirm the identities of the lipid species contributing to this variation by obtaining structural information using tandem MS (LC-MS/MS).
Evaluation of circulating biomarkers of bone metastasis in early-stage breast cancer.
Authors: Dean-Colomb Windy Marie , Hess Kenneth R. , Young Elliana J. , Gornet Terrie , Handy Beverly Carol , Moulder Stacy L. , Ibrahim Nuhad K. , Pusztai Lajos , Booser Daniel J. , Valero Vicente , Hortobagyi Gabriel N. , Esteva Francisco J.
Published: March 2020

10613

Background: Bone is the preferred site for metastasis of breast cancer, affecting approximately 70% of women with advanced disease. N-terminal of procollagen type 1 (P1NP), c-terminal peptide crosslinks (CTX), Osteocalcin (OC) and interleukin-6 (IL-6) are markers of bone turnover that may have clinical utility as predictors of breast cancer recurrence in the bone. Methods: Serum was collected from 168 patients with stage I/II/III breast cancer prior to treatment from 09/2001 to 12/2008 and stored at -80 C. Serum levels of P1NP, CTX, OC and IL-6 were determined using the Roche’s Elecsys 2010 automated immunoassay system. Correlations of biomarker levels with time to bone metastasis (BM) development were assessed with Cox proportional hazards regression analysis and the Kaplan-Meier method. Results: Among the 168 patients analyzed, 60 patients subsequently developed BM. The biomarkers all had skewed distributions with long right tails and thus were all analyzed on the log scale. Residual analysis suggested non-linear relationships between each biomarker and risk of developing BM during follow-up. Thus, we fit Cox proportional hazards regression models for each biomarker with quadratic polynomials (on the log scale). On univariate analysis, these analyses generated p = 0.33 for IL-6, 0.26 for osteocalcin, 0.40 for CTX, and 0.032 for P1NP. Adjusting for clinical factors (stage, age, race, post menopausal, ER/PR status, HER2 status, nuclear grade) yielded p = 0.0035 for the quadratic polynomial for log P1NP. A cut-point of 75 ng/mL identified patients with a short time to development of BM. The 1, 3, and 5-year freedom-from-BM probabilities were 96%, 77% and 66% for the 150 patients with P1NP values ≤ 75 ng/mL and 88%, 45%, and 36% for the 16 patients with P1NP values > 75 ng/mL. The hazard ratio comparing patients with P1NP values ≤ 75 ng/mL to patients with P1NP values > 75 ng/mL was 3.0 (95% CI, 1.5 - 6.2) and p = 0.0075 ng/mL. After adjustment for clinical factors, the hazard ratio was 3.4 (1.5, 7.6) with p = 0.0026. Conclusions: Serum P1NP levels >75 ng/mL correlate with a shorter time to development of BM in patients with stage I-III breast cancer.
Predictive impact of RRM1 protein expression on vinorelbine efficacy in NSCLC patients randomized in a chemotherapy phase III trial.
Authors: Vilmar Adam , Santoni-Rugiu Eric , Sorensen Jens Benn
Published: March 2020

10617

Background: Platinum based doublets remain the cornerstone of treatment in non-small cell lung cancer (NSCLC) and often includes gemcitabine. A biomarker predicting sensitivity to this antimetabolite would represent a major step forward in the management of advanced disease. Ribonucleotide reductase subunit M1 (RRM1) has been of some value in NSCLC but evidence is not uniform. Accordingly, we explored the predictive role of RRM1 in patients with advanced NSCLC. Methods: 443 patients with advanced NSCLC were randomized in a phase III trial to regimen A (paclitaxel and cisplatin with gemcitabine) or regimen B (cisplatin and vinorelbine). Immunohistochemical evaluation of RRM1 was performed on mainly bioptic material and correlated to response rates, progression free survival (PFS) and overall survival (OS). Results: 261 (58.9%) patients had representative tissue samples for RRM1 evaluation. Disease control rate, PFS and OS were significantly improved in patients with RRM-negative tumors receiving regimen B as compared to patients with RRM-positive tumors (68.8% vs. 31.2%, P = 0.046 , 6.90 months (mths) (95% CI 5.83-7.96) vs. 3.93 mths (95% CI 3.11-4.76), P = 0.000 and 11.57 mths (95% CI 9.65-13.48) vs. 7.4 mths (95% CI 5.37-9.43), P = 0.002, respectively). However, this was not the case for patients receiving regimen A (67.6% vs. 32.4%, P = 0.366, 6.73 mths vs. 7.6 mths, P = 0.207, 11.07 mths vs. 12.37 mths, P = 0.103, respectively ). Together with histological subtype, RRM1-positivity emerged as the only other significant prognostic variable with a hazard ratio of 1.56 (95% CI 1.38-2.35, P = 0.033) in multivariate analysis for patients treated with regimen B. Conclusions: RRM1 protein expression was without predictive impact in patients treated with cisplatin, paclitaxel and gemcitabine. Surprisingly, the predictive power proved robust in the cisplatin and vinorelbine arm across classic endpoints confirmed by multivariate analysis. These findings may suggest that RRM1 is involved in vinorelbine sensitivity and warrant further research which will be presented shortly.
Synthetic lethality of EGFR blockade in the context of tumor hypoxia.
Authors: Makino Haruhiko , Javvadi Prashanthi , Tomimatsu Nozomi , Saha Debabrata , Burma Sandeep , Nirodi Chaitanya
Published: March 2020

10624

Background: Hypoxia induces a wide spectrum of biological responses in tumors that alter tumor radio-sensitivity. In several tumors, including non-small cell lung carcinoma (NSCLC), the epidermal growth factor receptor (EGFR) is frequently over-expressed and activated in response to hypoxia, although the precise role of EGFR in hypoxia-associated radioresitance is unclear. Activating mutations in EGFR have been clinically linked to dramatic responses to EGFR tyrosine kinase inhibitors. We previously demonstrated that NSCLCs harboring activating EGFR mutations are radiosensitive with severe deficits in the non homologous end-joining (NHEJ) repair of DNA damage. Methods: We investigated hypoxia associated radiation responses in a panel of NSCLC cell lines and immortalized human bronchial epithelial cells expressing wild type EGFR (WT) or expressed EGFR forms with somatic activating L858R or DE746-E750 mutations (MT). Results: (1) Relative to WT EGFR, MT EGFR expression is associated with significantly higher radiosensitivity under chronic hypoxia (2) Under hypoxic conditions, MT EGFR expression is associated with a unique spectrum of DNA damage responses (DDR) that significantly deviated from canonical response of WT EGFR expressing NSCLCs (3) Genome wide gene expression analysis identified a dramatic hypoxia-associated DDR shift to selective down-regulation of homologous recombination (HR). Moreover, MT EGFR deficits in NHEJ caused a synthetic lethality effect in the context of hypoxia through a simultaneous blockade of HR and NHEJ. (4) Blockade of WT EGFR by the anti-EGFR monoclonal antibody, cetuximab perfectly recapitulated the contextual synthetic lethality of MT EGFR expression. (5) Cetuximab had a dramatic synergistic effect on survival of rat lung orthotopic tumors where hypo-fractionated radiotherapy or cetuximab alone had marginally controlled tumors. Conclusions: The data support a critical role for EGFR-mediated DNA repair in hypoxia associated tumor radio-resistance. Furthermore, this role could be a target for refractory hypoxic EGFR-WT tumor especially in the context of hypo-fractionated radiotherapy which, in its current form, is potentially counter-productive for hypoxic tumors.