bluuebunny/crossref_metadata_2025 huggingface dataset:
Internal mammary node biopsy for breast cancer patients: Issues for discussion and our practice.
Authors: He Qingqing
Published: March 2020

e22089

Background: The aims of this study was to determine the clinical implications of internal mammary node biopsy as staging, treatment with radiotherapy and systemic treatment and a prognostic factor in patients with breast cancer. Methods: Internal mammary node biopsy via intercostal space was performed in 344 cases of breast cancer. Anatomical location of internal mammary nodes were recorded. Pathological status of internal mammary node were detected by H and E stains. Results: Internal mammary node biopsy was successfully finished in 344 patients.There were 162 cases (48.26%) with positive axillary nodes, while the internal mammary nodes were involved in 72 cases (20.93%).53 patients (32.72%) had regional metastases in both the axillary and internal mammary lymph nodes.19 (5.52%) patients had internal mammary node metastasis but no axillary node metastases. PN stage migration was seen in 72 patients with a positive internal mammary node. There was no statistic relation between internal mammary nodes metastases and tumor location (X2 =0.48, P>0.05). There was no complication such as pneumothorax or haemorrhagia. Conclusions: The approach used is a reliable surgical technique for removing lymph node from intercostal space. Without exploring internal mammary nodes status, pN stage was incomplete. Internal mammary node biopsy enables treatment to be better adjusted to the needs of the individual patient. Using internal mammary node biopsy, patients with a negative internal mammary node can be prevented from radiation to internal mammary nodal areas. This leads to optimization of treatment.
An analysis of preoperatively HE4 and CA125 in matched samples of patients (pts) with benign disease and borderline ovarian tumors (BOT) with and without invasive implants (InvImp).
Authors: Nassir Mani , Braicu Elena Ioana , Richter Rolf , Chekerov Radoslav , van Gorp Toon , Timmerman Dirk , Vergote Ignace , Sehouli Jalid
Published: March 2020

e22102

Background: Presence of InvImp, quality of surgical staging and residual tumor mass were shown to be major prognostic factors for BOT. Currently, there are no biomarkers that can predict the various types of BOT. Aim of our study was to assess the value of individual CA125 and HE4 values, and the combination of CA125 and HE4 according to the ROMA algorithm for detecting BOT, and for predicting the presence of InvImp. Methods: BOT pts with InvImp (n=14) were matched by age and histology and compared to consecutive pts with benign disease and BOT without InvImp (1 to 4 to 1, respectively). Blood was collected prior surgery. HE4 and CA125 were determined using ELISA. The performance of HE4 and CA125 were analyzed using the receiver operator characteristic (ROC) and logistic regression. Results: HE4 and CA125 were increased in BOT with InvImp (median=73.85pM and 49.5IU/dl) compared with benign diseases and BOT without InvImp (median values: 48pM and 18.5IU/dl, and 61.45pM and 26.5IU/dl, respectively). HE4 and CA125 expression were significantly increased in BOT with InvImp compared to benign diseases (p=0.029 and 0.004, respectively) but not to BOT without InvImp. Furthermore we grouped BOT without InvImp together with benign diseases and compared them with BOT with InvImp. HE4 and CA125 cut off values of 65pM and 60IU/dl, respectively were independent predictive factors for the presence of InvImp (p=0.018, OR=5.32, 95%CI 1.33-21.20 and p=0.031, OR=4.83, 95%CI=1.16-20.13, respectively). ROMA performed better than HE4 or CA125 alone (p=0.002, RR=9.17, 95%CI=2.31-36.36). The OR of a high ROMA score were 0.082 (95%CI: 0.02-0.34; p=0.001) for benign diseases and 0.27 (95%CI: 0.05-1.42; p=0.123) for BOT without InvImp compared to BOT with InvImp. Conclusions: The ROMA score can not predict sufficiently InvImp in BOT. Further multicenter and larger prospective studies are needed.
Use of nanocurcumin to inhibit proliferation and metastases of hepatocellular carcinoma via NF-κB mediated matrix metalloproteinase-9 downregulation.
Authors: Xu Yang , Hu Bo , Anders Robert , Maitra Anirban , Fan Jia
Published: March 2020

e22103

Background: Curcumin regulates the expression and secretion of MMPs, and has potent anti-cancer properties in several human cancer cell lines and animal carcinogenesis models. In this study we try to investigate the effects of polymeric nanoparticle encapsulated formulation of curcumin– nanocurcumin (NC) on Hepatocellular carcinoma (HCC) in vitro and in vivo. Methods: The effects of NC alone and in combination with sorafenib (SO) were investigated on HCC cell lines, Huh7 and MHCCLM3 in vitro by using proliferation and invasion assay, western blot, qRT-PCR, enzyme-linked immunosorbent assay and immunohistochemistry staining, and the subcutaneous and orthotopic HCC xenograft nude mice models (MHCCLM3) were used to evaluate primary tumor growth and metastasis after treatments. Results: NC alone (or combined with SO)inhibited HCC cell proliferation (p<0.05) and invasion in vitro (p<0.01), and remarkably decreased both the subcutaneous and orthotopic primary tumor growth and lung metastases in vivo. NC and/or SO could down-regulating the expressions of Matrix Metalloproteinase-9 (MMP9), p-ERK1 and NF-kB/p65 (p<0.05). Conclusions: NC showed potent anti-invasion and metastasis properties in HCC via NF-κB mediated MMP9 down-regulation, which may provide a new strategy to the treatment of HCC patients and prevention of tumor recurrence after operation.
Capturing and molecular analysis of tumor cells isolated from ascites fluid to predict response to treatment.
Authors: Krahn Thomas , Jeffers Michael , Von Hoff Daniel D. , Seetharam Mahesh , Liu Xinjun , Kim Phillip Sangwook , Singh Sharat
Published: March 2020

e22110

Background: As access to tumor tissue if often limited in clinical studies, we are exploring technologies to capture tumor cells from body fluids with the aim to use the profile of pathway activation to predict response to treatment in early clinical trials. Methods: Tumor cells were captured from ascites fluid collected from an ovarian cancer patient via immunomagnetic enrichment. Isolated tumor cells were characterized for panel of signal transduction proteins at base line and after the treatment with PI3K inhibitor and/or MEK inhibitor alone or in combination. Comprehensive functional pathway modulation was determined using multiplexed Collaborative Enzyme Enhanced Reactive-immunoassay (CEER) to evaluate the treatment efficacy. Results: Sufficient numbers of tumor cells were present in ascites fluid for immunomagnetic enrichment and for subsequent functional pathway analysis. Comprehensive pathway modulations (HER1, HER2, HER3, cMET, IGF1R, PDK1, PI3K, AKT, PRAS40, RPS6, ERK, MEK and RSK) were determined with tumor cells using CEER. Pathway modulation upon in vitro drug treatment revealed insufficient targets as well as downstream signal protein inhibition against single agent PI3Ki or MEKi treatment. When these cells were treated with both compounds however, a dramatic target/pathway inhibition was observed. Conclusions: Viable, stimulatable tumor cells can be isolated in high number from ascites fluid and routine pathway analysis can be achieved in clinical setting. As pathway redundancy is a common feature in most solid tumor, a comprehensive in vitro analysis of pathway against various targeted drugs using tumor cells isolated from easily obtainable biological fluid (ascites, plural fluid, etc.) may guide clinicians in selecting the most effective targeted drug combination. Significant differences can be detected between single and combination treatment. Further studies need to be done to validate these findings.
Impact of antiangiogenic treatments on exhausted PD-1+ T lymphocytes in colorectal cancer.
Authors: Pernot Simon , Terme Magali , Marcheteau Elie , Voron Thibault , Colussi Orianne , Tartour Eric , Taïeb Julien
Published: March 2020

e22121

Background: Tumors develop immunosuppressive mechanisms to escape the immune system. Among these mechanisms, T lymphocytes can express inhibitory molecules such as Program Death-1 (PD-1) protein which impair their activation and their effector functions. Multi-target anti-angiogenic tyrosine kinase inhibitors (TKI) that are routinely used as first- or second-line treatment of cancer patients, have been shown to modulate immunosuppressive mechanisms especially regulatory T cells. However, the role of specific VEGF/VEGFR blockade on PD-1 expression by T lymphocytes has not been studied. Methods: PD-1 expression on T lymphocytes has been analyzed by flow cytometry in a mouse model of colorectal cancer (CT26) treated by anti-angiogenic molecules targeting the VEGF-A/VEGFR axis and in the peripheral blood of metastatic colorectal cancer patients. Results: Sunitinib that directly targets VEGFR, PDGFR, c-kit, but also anti-VEGF-A antibody (the mouse orthologue of bevacizumab) decrease PD-1 expression on T lymphocytes infiltrating the tumors. Interestingly, anti-angiogenic treatments decrease the percentage of lymphocytes expressing not only PD-1 but also Tim-3 suggesting that the most exhausted T cells are targeted by anti-angiogenic treatments. Administration of masitinib, a TKI acting on KIT, PDGFR and FAK but not on the VEGF/VEGF-R pathway, was not able to modulate PD-1 expression on T cells. Finally, anti-VEGF-A treatment (bevacizumab) associated with chemotherapy decreases the proportion of PD-1+ CD4+T cells in the peripheral blood of metastatic colorectal cancer patients. Conclusions: These results show that anti-angiogenic treatments targeting VEGFA/VEGFR decrease PD-1-expressing T lymphocytes in colorectal cancer. These results suggest that VEGF-A could be involved in PD-1 expression and maybe in the induction of T lymphocyte exhaustion in colorectal cancer. We and others have shown that anti-angiogenic molecules could modulate other immunosuppressive populations such as regulatory T cells and myeloid suppressive cells, anti-angiogenic molecules might be efficiently associated with immunotherapeutic strategies in colorectal cancer.
A comparative proteomic approach for identification of putative biomarkers of neoadjuvant chemotherapy resistance in breast cancer.
Authors: Hussain Tasadooq , Scaife Lucy , Hodgkinson Victoria , Agarwal Vijay , Mahapatra Tapan , Kneeshaw Peter John , McManus Penelope Louis , Lind Michael , Drew Philip , Cawkwell Lynn
Published: March 2020

e22123

Background: Neoadjuvant chemotherapy is a standard treatment for locally advanced breast cancer however chemoresistance can be a major obstacle in ER+ cancers. Using comparative proteomic approaches (antibody microarray/AbMA and 2D-PAGE with MALDI-TOF/TOF MS) to investigate a pilot series of breast cancer samples we recently identified 14-3-3 theta/tau, tBID and BcL-XL as putative biomarkers of response to neoadjuvant chemotherapy (Hodgkinson et al. J Prot 2012, 75:1276-1283 and 75:2745-2752). Here we aimed to analyse further samples using the AbMA approach and to re-analyse the combined data. Methods: Samples from chemoresistant and chemosensitive breast cancers were selected following anthracycline-taxane chemotherapy and 4 experiments were performed using ductal ER+ tumours. Differential protein expression was compared between chemoresistant and chemosensitive samples using the Panorama XPRESS Profiler725 AbMA kit. The combined data from 9 AbMA assays and 3 2D-PAGE/MS experiments was then analysed using Ingenuity Pathway Analysis (IPA; Ingenuity Systems). Results: 89 differentially expressed proteins (DEPs) were seen in the 4 further AbMA experiments. Sixteen proteins (DR4, E2F6, ILK, MeCP2, MyD88, PI19INK4d, PRMT2, ID, FANCD2, FAKpTyr577, Pinin, Zyxin, 14-3-3 theta/tau, BcL-xL, Pancytokeratin and RIP) were found in at least 2/4 experiments. In the combined dataset (12 experiments from 2 proteomic platforms), 8 DEPs were seen in at least 3 experiments. These were 14-3-3 theta, 14-3-3 epsilon, 14-3-3 gamma, Bcl-xl, Bid, Phosphokinase B, Vimentin and FAK. 121 DEPs from the combined data were analysed using IPA; 12 DEPs were mapped onto the PI3K/AKT pathway. Conclusions: We have identified at least 8 proteins which could play a role in chemoresistance and clinical validation of these putative biomarkers is currently underway.
Meta-analysis of cancer cell lines genomes based on their response to ttfields.
Authors: Giladi Moshe , Wasserman Yoram , Schneiderman Rosa S. , Porat Yaara , Munster Mijal , Weinberg Uri , Kirson Eilon David , Palti Yoram
Published: March 2020

e22125

Background: Tumor Treating Fields (TTFields) therapy is an established anti-mitotic treatment modality. The inhibitory effect of TTFields was demonstrated in numerous cancer cell lines with some cell lines exhibiting higher treatment sensitivity than others. The goal of the present study is to compare genomic characteristics of cell lines based on their response pattern to TTFields. Methods: Fifteen different human cancerous cell lines were treated using TTFields at their optimal frequency with the same nominal intensity (1.8 V/cm). The distribution of the response to TTFields was found to be bimodal with inhibitions of 30-40% and 50-60% compared to controls. Cell lines were divided into regular and high sensitivity groups according to their response to TTFields. Both groups were characterized based upon the distribution of mutated genes using the Cancer Cell Line Encyclopedia (CCLE) database. Results: Based on the distribution of mutated genes we identified 1 gene with significantly higher probability (P=0.017) to be present in the regular than the high sensitivity group. This gene (NEK3) is a never-in-mitosis (NIMA) related kinase 3 responsible for deacetylation of microtubules. In addition, 2 genes were found with significantly higher probability (P<0.05) to be present in the high- than the regular sensitivity group; ASPH, an aspartate beta-hydroxylase involved in negative regulation of cell proliferation through modulation of calcium homeostasis and TEC, a protein tyrosine kinase involved in immunological signaling pathways. Conclusions: The possible involvement of NEK3 mutations in determining sensitivity to TTFields is interesting in view of the known interference of TTFields with spindle microtubule assembly. ASPH mutations could release cells from negative regulation of proliferation, increasing their replication rate, and thus increasing their sensitivity to TTFields. In order to further test this hypothesis, a comparison of the effect of TTFields needs to be performed on wild type and knockout cell lines for these genes. A comparison of the mutation of these genes in tissue samples from patients whose tumors show high sensitivity to TTFields, may lead to a personalized approach to the use of TTFields clinically.
Ratios between VEGF ligands and receptors in tumor and stroma have impact on the outcome in resectable NSCLC.
Authors: Jantus-Lewintre Eloisa , Usó Marta , Sanmartin Elena , Gallach Sandra , Sirera Rafael , Blasco Ana , Hernando Cristina , Martinez Nieves , Figueroa Santiago , Casimiro Enrique , Camps Carlos
Published: March 2020

e22147

Background: In tumor angiogenesis there is a complex interplay between endothelial, stromal and tumor cells. Some key regulators of this process are the members of the vascular endothelial growth factor (VEGF) family of ligands and receptors and the neuropilins (NRP). This study analyzes the correlations between the expression of these angiogenic factors in tumor cells and tumor stroma, and their prognostic role in tissue samples from resected non-small cell lung cancer (NSCLC) patients. Methods: Representative tumor and stroma areas from FFPE tissue samples of 125 early-stage NSCLC patients were carefully micro-dissected. RNA isolated from the samples was retrotranscripted and preamplified. RTqPCR was performed using hydrolysis probes (TaqMan, Applied Biosystems) and relative quantification was calculated using GAPDH and CDKN1B as endogenous controls. Results were normalized against a human cDNA (Clontech) as a reference. All statistical analyses were considered significant at p<0.05. Results: Paired Wilcoxon test revealed differences between tumor and stroma gene expression for VEGFB (p<0.0001), VEGFC (p<0.0001), VEGFR-1 (p<0.0001), VEGFR-2 (p=0.020), VEGFR-3 (p< 0.0001), NRP-1 (p=0.001) and NRP-2 (p< 0.0001). Survival analyses showed that those patients with higher levels of the Ratio Stromal-VEGFA/ Tumoral-VEGFR-2 had worse time to progression (TTP) (median 26.23 months vs NR, p=0.013) and overall survival (OS) (median 29.50 months vs NR, p=0.001). Similarly, those patients with higher levels of the Ratio Stromal-VEGFC/Tumoral-VEGFR-3 had worse TTP (median 23.30 vs 70.53 months, p=0.015) and OS (median 37.20 months vs NR, p=0.023). Conclusions: Our results show significant differences in the expression of VEGF family members between tumor and stromal cells. This may indicate the importance of the tumor-stroma interaction when trying to understand the angiogenic process. Furthermore, the combination of the ligands expression in stroma and their receptors in tumor may have a prognostic value in NSCLC patients. Supported by grants PS09-01149 and RD06/0020/1024 from ISCIII.
Genetic variation in CYP19A1 and occurrence of adverse events in exemestane treatment with early breast cancer patients in the Dutch TEAM trial.
Authors: Houtsma Danny , Fontein Duveken , van der Straaten Tahar , Baak-Pablo Renee , Wessels Judith A.M. , Seynaeve Caroline , Van De Velde Cornelis J. H. , Nortier J. W. R. , Gelderblom Hans , Guchelaar Henk-jan
Published: March 2020

e22152

Background: Aromatase inhibitors (AI) are an important part of treatment of endocrine sensitive breast cancer. Adverse events in patients treated with AI’s often cause treatment discontinuation. The most common adverse events, such as arthralgia, myalgia and hot flushes are probably caused by estrogen deprivation and predict treatment efficacy. It is unclear which patients are at risk to develop these adverse events. The aim of this study was to examine if SNP’s in the CYP19A1 gene can predict the occurrence of adverse events in breast cancer patients treated with adjuvant exemestane. Methods: Patients of whom tissue was available and were selected from the Tamoxifen Exemestane Adjuvant Multinational (TEAM) trial. DNA was isolated from tumor samples and 30 SNPs were identified using a tagging SNP approach, aiming for 80% coverage of CYP19A1. Genotypes were determined with taqman assays. Primary endpoint of the study was the occurrence of adverse events. Secondary endpoints were the occurrence of hot flushes, arthralgia, and myalgia. Results: 807 patients were included in the analyses and genotypes were obtained in 722 cases. One SNP, rs8031311, was associated with a higher incidence of adverse events with an odds ratio of 2.8. Four SNP's were associated with an increased incidence of hot flushes: rs934635, rs4775928, rs16964189, rs6493496 with odds ratio’s of 2.9, 1.8, 1.8 and 2.6 respectively. No association was found between variation in CYP19A1 and the occurrence of arthralgia or myalgia. Conclusions: Germline variation in the CYP19A1 gene is related to the occurrence of adverse events, specifically hot flushes, in early breast cancer patients treated with exemestane. These findings may contribute to the individualization of hormonal therapy in breast cancer. [Table: see text]
Association between the founder brazilian TP53 p.R337H mutation and HER2-positive status.
Authors: Giacomazzi Juliana , Graudenz Marcia S. , Osorio Cynthia ABT , Koehler-Santos Patricia , Palmero Edenir Inez , Michelli Rodrigo Augusto Depieri , Fernandes Gabriela Carvalho , Scapulatempo Neto Cristovam , Achatz Maria Isabel Waddington , Martel-Planche Ghyslaine , Soares Fernando A , Caleffi Maira , Goldim Jose Roberto , Hainaut Pierre , Camey Suzi Alves , Ashton-Prolla Patricia
Published: March 2020

e22154

Background: Recent studies in North American and British series have suggested an association between germline TP53 mutations and early onset HER2-positive breast cancer (BC). Mutations in the TP53 gene are estimated to occur in 1:2,000-1:5,000 individuals of the general population. Among women with BC who are unselected for family history they occur in up to 0.25%. In women with early-onset BC (<30 years) they occur in up to 7%. A specific germline TP53 mutation (c.1010G>A; p.R337H) has been encountered in 0.3% of the general population, in 13.3% of Li-Fraumeni-like families and in 0.5-2.4% of BC-affected women in Brazil. In an exploratory approach, the aim of this study was to investigate, in our series, if there is an association between human epidermal growth factor receptor 2 (HER2) amplification and the germline TP53 p.R337H mutation in a series of BC-affected women. Methods: A series of 718 Brazilian BC-affected women was genotyped in our previous study and 64 (8.9%) were mutation carriers. These cases were recruited from 3 centers and HER2 analysis was performed by immunohistochemistry (IHC) in each center according to validated protocols. Statistical analysis was done using SPSS software. Results: In 47 of 64 mutation carriers (73.5%), BC showed HER expression (either 1+, 2+ or 3+) while among 654 non-carriers, this immunophenotype was observed in 320 (48.9%) (p<0.001). This pattern was still observed after stratification of groups according to age at diagnosis (≤45 and ≥55 years) (p<0.001 and 0.03, respectively) and was independent of recruiting center. Conclusions: These results indicate that BC developing on a background of the founder Brazilian TP53 p.R337H mutation shows more frequently some amplification of HER2, consistent with recent studies showing an association between germline TP53 mutations and BC with HER2 hyperexpression. These findings should be confirmed in larger series to understand the significance of the association. If confirmed, this finding may justify germline TP53 p.R337H mutation testing in all HER2-positive Brazilian BC patients and would have a significant impact on patient and family counseling as well as on treatment.
A score based in toll-like receptors expression to predict prognostic in molecular subtypes of breast cancer treated with neoadjuvant chemotherapy (NAC).
Authors: Pinto Joseph A. , Flores Claudio J. , Valdiviezo Priscila I. , Velazco Rodolfo , Garay MIguel , Suazo Juan F. , Aguilar Alfredo , Gomez Henry , Vallejos Carlos S.
Published: March 2020

e22165

Background: To evaluate toll-like receptors (TLRs1-9) and IRAK 1,3 and 4 expression in a breast cancer patients dataset that were treated with NAC as prognostic factors in terms of recurrence-free survival (RFS). Methods: Weretrieved normalized gene expression data from a public database (GEO: GSE25066) that includes 253 samples of breast cancer patients treated with NAC (antracyclines/taxanes) profiled with the U133A microarray. The median of RFS was 3.2 years (52 relapses observed). Distribution of molecular subtypes was: Luminal A (31.2%), Luminal B (17.4%), HER2 (7.1%), Basal (36.4%) and Normal (7.9%). We build a ratio of target gene/reference gen (ATCB) with to obtain replicable cut-points in other datasets. Levels of Gene expression were divided in three cut-points (Q1, Q2, and Q3). A cox-model was used to select gene expression cut-points with significant effects in the RFS. Results: According to molecular subtypes, there were significant differences in expression of TLR5 (higher in basal and Luminal A), TLR6 and IRAK1 (higher in basal and HER2). There were not differences of expression according to the pathologic response. The multivariate analysis shown that genes associated with the RFS were: TLR5 (HR=2.1 for the lower quartile), TLR6 (HR=2.4 for upper Q2), IRAK1 (HR=2.0 for upper quartile) and IRAK 3 (HR=2.2 for upper Q2). A TLR-Score was constructed (1 point added for each unfavorable gene-group). Cases were grouped in two categories (score 0-1 as good prognostic and score ≥ 2 as poor prognostic). There were and overall significant differences in both score-groups (poor prognostic: HR=4.9). In Basal and luminal B tumors there were significant differences (logrank test: P<0.001 and P=0.014, respectively), in HER2 tumors there were differences although not significant (P=0.143). There were not differences in Luminal A tumors. Conclusions: These data support the exploration of a TLR expression-based score to predict for RFS in breast cancer treated with NAC.
Feasibility study of SNPs detection associated with breast cancer by genome-wide association virtual studies.
Authors: Kooshyar Mohammad Mehdi , Nassiri Mohammadreza , Aslaminezad Aliasghar , Betaraf Morteza
Published: March 2020

e22167

Background: Available genomic data and genome-wide association virtual studies (GWAS), provide possibility of genetic markers detection known to be associated with the complex diseases. Genome-wide association studies, involving direct testing of genetic polymorphisms in large series of disease cases versus controls, provide a powerful approach to identify lower penetrance alleles that cannot be detected by genetic linkage studies. Methods: By utilizing genotyping platforms that can type hundreds of thousands of SNPs simultaneously, it is possible to conduct association studies using sets of SNPs that tag most known common variants in the genome, and hence scan associations without prior knowledge of function or position. GWAS have been conducted in five of the most common cancer types: breast, prostate, colorectal, lung and melanoma. GWAS of breast cancer was performed by simulation of 7 known SNPs on chromosomes 2, 5, 6, 8, 10, 11 and 16 for 10,000 women. Results: The strongest associations were found for rs2981582 in the FGFR2 gene. SNPs: rs889312 ° rs2180341 and rs3817198 were associated with breast cancer in benferroni significant level. However, SNPs: rs 13387042 ° rs2180341 and rs13281615 on chromosomes 2, 6 and 8 were not associated with breast. Conclusions: Results of this research show that the detection of SNPs associated with disease is easily possible through employing virtual systems based on real data of Hap Map project by using R software.
Galectin-3: A possible complementary marker to the PSA blood test.
Authors: Raz Avraham , Balan Vitaly , Wang Yi , Nangia-Makker Pratima , Kho Dhonghyo , Heilbrun Lance K. , Bajaj Madhuri , Smith Daryn W. , Heath Elisabeth I.
Published: March 2020

e22168

Background: The prostate-specific antigen (PSA) is a test widely used to detect, monitor, and predict the biology of prostate cancer (PCa). Galectin-3 (Gal-3) is a carbohydrate-binding protein and is present in the seminal fluid. It is a tumor-associated protein, which plays a role in metastasis, apoptosis, and tumor growth. We recently reported that PSA cleaves Gal-3. Cleavage status of Gal-3 in the prostate tissue might be associated with the tumor formation and progression of PCa. Therefore, we compared Gal-3 levels in the blood of non-cancer urology patients and metastatic PCa patients. Methods: Non-cancer and metastatic PCa patients were recruited from the genitourinary oncology clinics. Sixteen men (8 PCa patients and 8 controls) were recruited during their clinic visit from May to August 2012. The metastatic PCa patients could be either castrate-sensitive or castrate-resistant. The control patients could not have any history of invasive cancer. A blood sample was collected from each study patient. Levels of Gal-3 were analyzed by Western blot and ELISA. The study design was a small, prospective, single institution pre-pilot study. The primary statistical endpoint was the mean level of serum galectin-3. Within each of the two groups of patients, it was desired to estimate the mean Gal-3 level to within 0.50 standard deviations (SD’s) of the true mean, with 80% confidence. This study design required N=8 men per group. Results: The cancer-free control patients had lower levels of serum Gal-3 than did any of the 8 PCa patients (see Table). These results suggest that serum Gal-3 levels in metastatic PCa patients and cancer-free controls are quite different. The serum PSA distributions of these 2 patient groups overlap slightly, hence PSA did not distinguish them quite as well as did Gal-3. Conclusions: This is the first report to show serum Gal-3concentrations were uniformly higher in patients with metastatic PCa as compared to non-cancer control patients. [Table: see text]
Observational study of real world clinical performance of microrna molecular profiling for cancer of unknown primary (CUP).
Authors: Sanden Mats Olot , Wassman Robert , Ashkenazi Karin , Benjamin Hila , Spector Yael , Goren Eran
Published: March 2020

e22173

Background: Molecular profiling of CUP may effectively identify the underlying tumor type and potentially influence treatment decisions and outcomes. A microRNA (miR) array has been shown in concordance studies at leading academic centers to perform well in correctly identifying the tumor of origin in CUP. This series of consecutive referrals tested in a CLIA-laboratory extends these findings to the real world clinical setting. Methods: MicroRNA was isolated from 258 consecutive specimens referred to Rosetta Genomics, analyzed on a 64 miR-based array, and interpreted for any of 42 specific tumor types as previously described. The laboratory medical director routinely collected follow-up information on correlation of test prediction with other clinical information, pathology findings, supplemental IHC, treatment response, and clinical course as well as physician perception of relevance and utility of test result. This information was scored categorically into 3 classes of concordance depending on agreement with clinical and/or pathological data. The composite data was de-identified and analyzed for assessment of overall test performance in this real world clinical setting. Results: Of 258 specimens submitted 90% were paraffin embedded tissue (FFPE). Sufficient tumor material was received in 217 (85%) cases, of which 192 (88%) were successfully processed and reported with a mean TAT of 7 days. The most common result was colorectal (12%), followed by breast (10.4%), upper body squamous cell (7.3%), ovarian (6.8%), and biliary tract/pancreatic (6.8%); and the remaining 109 cases (57%) represented 30 additional tumor types. Overall the concordance with the clinical and/or pathological best final diagnosis was 86%, and when the reporting algorithm yielded a single diagnosis (51% of cases) the concordance was 89%. Clinical utility as measured by consideration of a shift in therapy or significantly increased decisional certainty for therapy was observed in approximately 70% of the cases. Conclusions: MicroRNA analysis in CUP cases performs comparably in the real world setting to that previously reported in academic studies and adds information of value to the management of patients.
Evaluating the prognostic significance of JAK2 in distant breast cancer recurrence.
Authors: Blau Carl Anthony , Miller Christopher P. , Kortum Amanda N. , Thorpe Jason D. , Schummer Michel , O'Briant Kathy , Coy Catherine M. , Beatty J David , Urban Nicole D.
Published: March 2020

e22178

Background: Estrogen, progesterone, and epidermal growth factor receptor ligands stimulate growth in a subset of breast cancers, however recent studies suggest roles for additional factors such as interleukins-6 and 8, prolactin, and erythropoietin. These and other growth factors act upon ligand-specific receptors to activate janus kinase 2 (JAK2), and JAK2 inhibitors are under investigation as a novel targeted therapy. We tested whether erythropoietin receptor (EPOR) or JAK2 mRNA levels are associated with distant breast cancer recurrence. Methods: We used quantitative RT-PCR to measure mRNA levels of JAK2, EPOR and a series of control genes using archival tumors from 112 women who experienced distant breast cancer recurrence (cases) and 112 tumors from women who did not (controls). Cases and controls were matched for tumor size, lymphovascular invasion, nodal status, extra-nodal extension, and ER/PR/HER2. Recurrence risks were evaluated using logistic regression. Associations between mRNA levels (via microarray data) and recurrence-free survival were validated in an independent cohort from the Netherlands Cancer Institute (n=295) using Cox proportional hazards regression. Results: Increasing JAK2 mRNA levels strongly correlated with a reduced risk of distant recurrence in our case control study (univariate p=0.0004, multivariate p=0.003), and with improved recurrence-free survival in the validation cohort (univariate p=0.0009; multivariate p=0.003). Remarkably, in the validation cohort, the ranking of the prognostic significance of JAK2 (top 3.5% of ~25,000 genes) was comparable to the known strong prognostic indicator of recurrence, ESR1 (top 1.3%). Similarly although less prominently, increasing EPORmRNA levels were significantly associated with reduced distant recurrence in our case control study (continuous model: univariate p=0.01, multivariate p=0.05) and with improved recurrence free survival in the validation cohort in univariate (p=0.03) but not multivariate analysis (p=0.35). Conclusions: JAK2 mRNA levels in breast tumors correlate with a reduced risk of recurrence. Understanding the mechanistic basis for this association is important for the rational application of JAK2 inhibitors.
Elimination of both cancer stem cells and progenitor cells as essential for tumor eradication.
Authors: Marak Creticus Petrov , Guddati Achuta Kumar
Published: March 2020

e22179

Background: Cancer stem cells (CSC) have been identified in several malignancies. Elimination of CSCs requires precise identification of markers and development of mechanisms to target them specifically. It is possible that in spite of elimination of CSCs, the tumor may recur. Dedifferentiation of progenitor cells may help replenish cancer stem cells. Methods: A mathematical model of a cancer stem cell system with four layers has been constructed to reflect the hierarchy of cancer stem cells, progenitor cells and, transit-amplifying cells and pre-differentiated cells. This model is constructed in the lines of the phase space model described by Kirkland. Dedifferentiation parameters were introduced to allow for dedifferentiation when the stem cell pool receded below 40% of its normal size. The effects of stem cell antagonists (SCA) and differentiating agents (DA) were simulated to study the perturbations caused by them on the growth of the tumor. Results: SCAs when used alone result in the establishment of a new equilibrium across different layers within the tumor by de-differentiation. DAs which act only on progenitor cells cause an increase in the number of cells in the subsequent layers and contribute to an increase in the size of the tumor. DAs which act only on the pre-differentiated layer of cells move cells from this layer into a terminally differentiated layer of cells which results in a new but lower equilibrium. DAs which act on the progenitor, transit amplifying and pre-differentiated cells cause the establishment of a new equilibrium which is lower than that of all agents which act on individual layers of cells. A combination of an SCA and a DA which acts on multiple layers eliminates the re-establishment of a new equilibrium and leads to decline in cell populations in all the layers, thus reducing the possibility of recurrence. Conclusions: SCAs when used individually are not effective in eliminating the tumor. DAs which act only on progenitor cells can fuel tumor growth but DAs which act on multiple layers have a profound effect in slowing the growth of the tumor but do not eliminate the tumor. This model implies that a combination of an SCA and a DA which acts on multiple layers may have a better chance at eliminating the tumor.
Triggers in the pathway from lung cell damage to malignancy: The glycans.
Authors: Saeed Anwaar Mohammed , Alkhazna Ammar Ali , Herndon Betty , Dim Daniel
Published: March 2020

e22189

Background: It has been reported that carbon nanoparticles (CN) produce mesothelioma as readily as do asbestos fibers. We have studied early CN damage in experimental animals, quantifying toxicity by release from cells of the nuclear chaperone HMGB1. Damage-released HMGB1 binds to receptors (RAGE, CD24) to upregulate cytokines and produce regulatory signals that control cell homeostasis. Further, HMGB1 has recently been shown to form a complex with P53, one of the most commonly mutated genes in human cancers, and HMGB1 is the controlling factor in autophagy, an important new field in translational cancer research. Methods: One HMGB1 receptor, CD24, is the source of a whole-receptor antibody used to determine extent of malignancy during surgery. CD24 is highly glycosylated. We hypothesized that carbohydrate vs protein specificity would show differences in tumor character. Would antibodies to the CD24 carbohydrate moiety reflect a tumor glycan character, and how would this relate to the tumor profile such as metastasis? With cooperation of TMC pathology Dept, blocks (N=15 each) of lung squamous cell, adenocarcinoma, and small cell cancer were sectioned and stained with mouse monoclonals to CD24, IgM –glycan, IgG-protein fraction. Results: Small cell and adenocarcinoma lung tumors showed mostly similar staining by CD24 protein vs CD24 glycan antibodies. Squamous cell CA showed elevated carbohydrate CD24 staining with negative or low protein stain in over half of the cases, and examination of patient records are being studied to determine metastatic character, lung- damage history and tumor chemotherapy response. Conclusions: Squamous cell tumors demonstrate significantly more staining by antibody to the glycan moiety of CD24, p=0.02. There was no relationship between numbers of positive nodes and staining, although other chart information awaits analysis. Research continues into the significance of this glycan CD24 receptor for HMGB1, our marker for nanoparticle lung damage, and its relationship to malignant transformation. [Table: see text]
The value of promoter methylation of the MGMT gene and IDH mutations in secondary glioblastomas.
Authors: Bie Li , Li Qian Yun
Published: March 2020

e22193

Background: Several studies have observed a change in MGMT silencing and IDH1 mutation in secondary glioblastomas (sGBMs). However, reports about the prognostic value of promoter methylation of the MGMT gene and IDH mutations in secondary glioblastomas (sGBMs) are few in number. Methods: The study involved primary and secondary tumor tissue samples from 89 GBMs pts (P/S: 42/47) diagnosed and treated within the First Hospital of Jilin University from Jan 2006 to Nov 2011. After surgical treatment, all GBM pts were subjected to radiotherapy with concomitant administration of TMZ. Pts with GBM were screened for promoter status of MGMT gene, by the methylation-speciﬁc polymerase chain reaction (MSP), and, for IDH mutations by direct sequencing. Results: A total of 42 of 89 pts (47.2%) had primary GBMs (pGBMs) (Group 1), while 47 pts (52.8%) had secondary GBMs (Group 2). In Group 2, 23 pts (21/47, 44.7%) developed a sGBM through progression from a low grade glioma (LGG) WHO grade II and a secondary anaplastic gliomas (sAG) (Group 2a), whereas 26 pts (26/47, 55.3%) showed a direct malignant transformation from a LGG to a sGBM (Group 2b). MGMT methylation was observed in 38 (42.7%) GBMs, with a higher frequency in sGBMs than pGBMs (24/14, 51.1 vs. 33.3%; p = 0.008). MGMT methylation status was associated with increased median survival time in pGBMs and sGBMs pts. IDH1 mutations are present in the majority of sGBMs but rare in pGBMs (31/5, 66.0 vs. 11.9%; p < 0.001). Group 2a had a higher frequency with IDH1 mutation than group 2b (18/12, 78.3 vs. 46.2%; p < 0.01). The median survival time after malignant progression of all sGBMs pts with an IDH mutation was longer than in pts with wild-type IDH (3.2 vs. 1.1ys; p < 0.01). IDH1 mutation had an improved outcome in group2a, which is compared with group 2b (3.9 ys vs. 2.1 ys; p < 0.05). Conclusions: In our population, pts with sGBM and IDH mutation had a signiﬁcantly improved outcome. In addition, MGMT methylation is also a powerful prognostic marker in sGBM pts.
Immune-phenotype assessment of ovarian cancer stem cells.
Authors: Martinez Maria Jose , Caballero Miguel , Martinez-Serrano Sergio , Vidal Laura , Fernandez Aranzazu , del Pino Marta , Vilella Ramon
Published: March 2020

e22200

Background: Current evidence suggests the presence of cancer stem cells (CSC) in epithelial ovarian cancer (EOC) and their role in chemoresistance and relapse. CD44+/CD24-, CD117+ and CD133+ expression have been reported as potential markers for CSC. Methods: Tumor cells from 5 patients with chemo-naive EOC and cells of ovarian and tubal epithelium from 5 healthy menopausal women undergoing surgery for benign pathology were cultured in standard medium (mediumA) and medium for embrionic cells (mediumB).Cells forming non-adherent spheres where considered CSC-like, and adherent cells as non-CSC-like. The percentages of CD24+, CD44+, CD117+, CD133+ and VEGF-R+ cells were determined by flow-cytometry. Results: After 1-7 week sphere formation was observed in all EOC-cells cultured in mediumB but in none of the healthy controls. Mean percentages of markers expression are shown below. Conclusions: Contrary to published data, expression of CD44+/CD24- was lower in CSClike; in Non-CSClike CD133+ was higher in EOC-cells compared to controls. Inmunophenotype of CSClike is still lacking and further investigation is warranted. [Table: see text] [Table: see text]
Immune checkpoint targets on circulating epithelial cells in early and late breast cancer.
Authors: Habets Leo , Frenken Bettina , El Ghali Ilham , Koerber Wolfgang , Danaei Mahmoud , Kusche Manfred , Kroll Torsten , Pachmann Katharina
Published: March 2020

e22201

Background: Our ongoing investigation of circulating epithelial cells(CEC) in early and late breast cancer with the MAINTRAC method (1ml blood, RBC lysis and fluorometric detection) has shown that CEC evade by hypoxia driven EMT (epithelial mesenchymal transition).We saw CEC with EMT and stemcellmarkers in early and late breast cancer. These cells can be found also in non cancer conditions. So far no differences in marker expression of CEC in cancer and noncancer conditions were found. This underlines the need for extended(e.g four color) analysis and allows searching for new promising targets like the immune checkpoint target PD-L1. Methods: We slightly adapted the basic MAINTRAC methodology to allow four color detection on the AMNIS FlowSight device.The IDEAS software allows comprehensive analysis of the populations. Beside four color analysis of single cells, TCA (tumor cell aggregates ) can be detected and characterized. The basic panel (EPCAMAF660.CD45PE,ALDH1FITC,DAPI) allows detection of living and death CEC and expression of the stemcell marker ALDH1. A second panel allows detection of other targets like PD-L1. Results: With the three color based original method the blood of 188 patients with early and 62 with late breast cancer were examined. No differences between conventional subgroups nor molecular subtype groups were found. In a large non cancer (n=256) control group we found high CEC counts in a group of patients with liver affections (mostly NAFLD). In these patients CEC showed the same marker ecpression .In late breast cancer only 60% of patients showed detctable cell counts.In patients with agressive disease (triple negative or HER2 enriched) mostly none or small CEC numbers were detectable.With our recently started (so far n=48) four color assay we detected coexpression of ALDH1 and PD-L1 which are more specific markers then CD44high or vimentin. Conclusions: Extended four color analysis on CEC allows monitoring of living CEC with stemcellmarker expression. Detection of immune checkpoint determinants like PD-L1on CEC is possible, further unraveling the biology of early and late breast cancer.
Abnormalities in cancer patients' coagulation system.
Authors: Gonzalez Florez Hector Alirio , Rojas M. , Quiroz J. , Mancera Y.
Published: March 2020

e22207

Background: It is well known cancer patients may present both hemorrhagic and thrombotic abnormalities and these phenomena are difficult to diagnose and treat. Thrombotic phenomena are developed clinicly in 15% in cancer patients and it’s higher in adenocarcinomas and myeloprolipherative disorders. These complications can be the consequence of coagulations factor direct activation or chronic DIC, because of the acquired capacity of neoplastic cells for starting coagulation cascade. Methods: In this study an untreated cancer patients group were compared with another healthy controls group, older than 14 years. Patients before treatment and controls were laboratory tested. A t – test for independent samples was employed. Results: Number: patients group: 66, control group: 73. Median age: patients: 50 (17-78). Controls: 52 (18 -93). Sex: Patients: men: 35%, women: 65%. Controls: men: 52%, women: 48%. Malignancies: breast cancer 9.7%; gastric carcinoma 8.1%; cervical carcinoma 6.5%; NH lymphomas 6.5%, unknown origen metastatic carcinoma 4.8%; oral carcinoma 4.8%. Clinical stages: I: 6.3%; II: 6.3%; III: 22.9%; IV: 62.5% (metastatic) 47.9%. Thrombotic antecedents in patients group: 19.7%. There were significant differences in: Platelets count: patients: 281121, controls: 307745 (p<0.001). Protein C: 0.87 and 3.81 UI/mL, respectively (p < 0.001). Antithrombin III: 97 and 101% (p<0.001). Plasminogen: 92.9 and 101.1% (p < 0.001). Lupus anticoagulant (RVV time): 27 and 25.6” (p<0.001). There were no significant differences in PT: 18.3 and 16.9” and PTT: 31 and 34”. The same significant differences were got when the patients of every clinical stage were compared with control patients. There were no differences when the patients of every clinical stage were compared among themselves, nor between sex. Conclusions: This study shows cancer patients may have many coagulation abnormalities: anticoagulant system defects and fibrinolytic system hipoactivity, demonstrated by the significant differences found in antithrombin III, protein C and plasminogen levels between patients and controls groups. It is suggested to practice these tests to cancer patients, even without clinical coagulation disorders, to stablish prophylactic measures.
Spontaneous and cyclophosphamide-enhanced metastasis inhibited by traditional Chinese medicine (TCM) herbal mixture LQ.
Authors: Zhang Lei , Wu Chengyu , Zhang Yong , Hoffman Robert M.
Published: March 2020

e22209

Background: Cyclophosphamide pretreatment of mice enhances metastatsis of HT1080 human fibrosarsoma cells (Cancer Res. 68, 516-520, 2008). Methods: HT1080 human fibrosarsoma cells were injected into the footpad or were injected into the epigastric cranialis vein of mice pre-treated with cyclophosphamide (CYC) or a combination of CYC and the herbal mixture LQ. The efficacy was monitored through dynamic subcellular imaging of trafficking of cancer cells in lymph nodes and blood vessels in live mice. Results: LQ significantly inhibited HT1080 spontaneous metastases to local lymph nodes in the foot-pad-injection model. In the control group, 100% of mice had lymph node metastasis, compared to 12.5% of the mice in the LQ treatment group. LQ also significantly inhibited CYC-enhanced experimental metastasis in the epigastric cranialis vein injection model. LQ prolonged the overall survival of mice in the spontaneous-lymph-node metastatic footpad-injection model. Conclusions: The TCM herbal mixture LQ shows promis to inhibit metastasis and will be further tested in other metastatic models.