bluuebunny/crossref_metadata_2025_split huggingface dataset:
Changes in phosphorylation and dephosphorylation status of cytoskeleton and their regulator proteins in CD34+ stem cells after G-CSF stimulation and in AML
Authors: Lannert H. , Lenze M. , Able T. , Park B. J. , Lenze A. , Meissner S. , Eckstein V. , Ho A. D. , Leicht S. , Franz T. 
Published: August 2020

e22067 Background: The cytoskeleton is a cellular ‘scaffolding‘ or ‘skeleton‘ contained within the cytoplasm. It is a dynamic structure that maintains cell shape, enables cellular motion. ADF/cofilin is a family of actin-binding proteins which disassembles actin filaments. LIMK2 is phosphorylated and activated by ROCK, a downstream effector of Rho, and phosphorylates cofilin, inhibiting its actin-depolymerizing activity. Profilin binds to actin. At high concentrations, profilin prevents the polymerization of actin, whereas it enhances it at low concentrations. Cdk1 phosphorylates beta tubulin. In mitotic cells, phosphorylated tubulin is excluded from microtubules, being present in the soluble tubulin fraction. Stathmin/Op18 is a cytosolic phosphoprotein which regulates the dynamics of microtubules. The process of tumor proliferation and metastasis is characterized by high rates of mitosis and migration into distant tissues. In this study we investigated the expression of cytoskeleton proteins in native hematopoietic CD34+ stem cells from BM in comparison to mobilized peripheral blood stem cells (mPBSCs) from G-CSF stimulated donors as well as CD34+ cells from AML. Methods: An Auto-MACS (Miltenyi) and FACS Vantage SE cell sorter (Becton Dickinson) was used to process high enriched (>99%) CD34+ cells fractions from MNCs. Proteomics was performed with the Proteome Works System (BioRad). Protein identification was carried out by Q-TOF and ion trap mass spectrometer analyses. Phosphoproteins were detected with ProQ-Diamond stain and specific antibodies. Results: The comparative proteome analyses show that 1. actin, tubulin, vimentin are overexpressed and phosphorylated, 2. cofilin and profilin are unphosphorylated, and 3. Stathmin is overexpressed in G-CSF mobilized hematopoietic stem cells and in AML in his active ‘dephosphorylated‘ form. Conclusions: Highly enriched hematopoietic CD34+ cells are a prerequisite for the qualitative and quantitative phospho/proteome analyses. Our results show, that mobilized stem cells “in vivo” and AML cells increase cytoskeleton proteins expression and cause a complex phosphorylation status, which may explain the regulation of migration and metastasis. No significant financial relationships to disclose.
The use of insulin and the effect on survival of non-small cell lung cancer patients
Authors: Farhan S. Y. , Jankowski M. , Hanbali A. , Wang D. 
Published: August 2020

e22073 Background: Diabetes mellitus (DM) is one of the common morbidities in United States. Insulin therapy has been frequently used in its treatment. Some studies have postulated that activation of Insulin and Insulin-like growth factor (IGF) pathway may contribute to cancer proliferation, growth and resistance to anticancer therapies. The impact of insulin on diabetic patients with non-small cell lung cancer (NSCLC) has not been reported. Methods: 1,233 patients with NSCLC diagnosed between January 1999 and December 2004 were identified from the Tumor Registry at Henry Ford Health System. Based on inclusion criteria, data from 1,206 patients were extracted from the electronic medical records. Statistical analyses were performed between insulin and non insulin users. Results: Out of 1,206 patients, 193 patients had NSCLC and DM. Data was available from 192 patients. Sixty (31.3%) were on insulin at the time of NSCLC diagnosis and defined as insulin users, while 132 were not on insulin and defined as non- users (68.7%). Demographics, co-morbidities, disease stage at diagnosis, therapeutic interventions, and laboratory values were analyzed for overall and cancer-specific survival. More insulin users (81%) than non-users (64%) presented with advanced disease (stage ≥ 3) at diagnosis (P=0.02). Although no significant difference of Hemoglobin A1c was observed, the median survival from all causes of death was 6.5 months for insulin-users versus 9.9 months for non-users (P=0.08). When survival time was calculated from cancer-specific death, the median survival between insulin and non-insulin users was 7.2 months versus 38.7 months, respectively (P=0.002), with a hazard ratio (HR) for insulin use equals 1.91. Results from our univariable and multivariable analyses will be presented at the ASCO annual meeting in detail. Conclusions: This data indicates that the use of insulin may adversely affect the biology and clinical course of NSCLC with a tendency of advanced disease stage at diagnosis and a shorter cancer-specific survival time. Prospective study of DM with NSCLC or other cancer diagnosis will improve our understanding of this potential adverse effect on cancer biology and provide scientific rational to target IGF-pathway especially in diabetic patients. No significant financial relationships to disclose.
A pilot study of biological markers (BM) as predictors for response and prognosis in patients (pts) with R0 resected gastric cancer (GC)
Authors: Li G. , Zhang Z. , Ma X. , Sheng W. , Zhu J. , Cai G. , Cai G. , Liu M. , Gu W. , Li J. 
Published: August 2020

e22076 Background: To evaluate clinical value of biological markers and their prognostic significance in gastric cancer as well as prediction to clinic outcome; To select preference marker for targeted therapy in high risk patients hence to guide personalized treatment. Methods: Retrieved from total 394 patients treated during the period of Jan 2004 to June 2006 of Cancer Hospital, Fudan University with complete medical records and follow up data, 84 patients with pre- and post- operative biological markers examined were selected. All of these patients underwent R0 gastrectomy, 61 (72.6%) patients underwent D2 resection. Forty- one (48.8%) patients received ≥ 4 cycles adjuvant chemotherapy. Preoperative tumor markers include: CEA, AFP, CA199, CA50, CA72–4, CA24–2 and postoperative immunohistochemical examination include: Bax, Bcl-2, P27, CyclinD1, TOPO2, MDR, GST-π, Ki67, AgNOR, EGFR, P21, P53, PCNA, C-myc and Neu were analyzed. Univariate, multivariate and Logistic regression analysis were performed and Kapan-Meier estimates were calculated. Results: For the whole group, 2-year OS and PFS were 76% and 66% respectively. Logistic regression analysis showed bcl-2, ki67, c-myc2 and Neu were relative with more nodal involvement. Patients with lower level of CEA, CA72–4 and higher level of cyclin D1 suivived longer than the other patients. There were different clinical significance with those biomarkers. Patients with lower level of cyclinD1, CEA had more local regional recurrence (p=0.026) while patients with higher PCNA experienced more distant metastasis (p=0.005). Univariate analysis showed that clinical staging, level of cyclinD1 and CEA were correlated with OS (p<0.05). Conclusions: Biological markers may be helpful in predicting the clinical outcome and prognosis for R0 resected gastric cancer patients. Further study are needed to confirm their clinical significance. No significant financial relationships to disclose.
Comparison of Lynch predictive models for identification of MLH1/MSH2 mutation carriers in a Spanish multicentre clinic- based cohort
Authors: Valenzuela I. , Balmaña J. , Rue M. , Blanco I. , Torres A. , Ramon y Cajal T. , Chirivella I. , Guillen C. , Brunet J. , Tejada I. 
Published: August 2020

e22077 Background: Different predictive models for Lynch syndrome have recently been developed and their comparative performance in a clinic-based cohort has not been assessed. We aimed to analyze the accuracy of the MMRpro, Barnetson, and PREMM1,2 models to predict MLH1/MSH2 mutation carrier status in 564 unrelated probands with clinical suspicion of hereditary colorectal cancer and compare it with Wijnen model and clinical criteria. Methods: Overall, 538 individuals (95%) underwent mismatch repair (MMR) deficiency screening before germline genetic testing (sequencing with or without large rearrangement analysis) and 26 (5%) performed direct genetic testing. Prediction scores for all individuals were calculated by each model. Sensitivity, specificity, positive predictive value (PPV), and the areas under the receiver operating characteristics curves (AUC) for all models were calculated and compared with the Revised Bethesda Guidelines (RBG). Results: 114 individuals (20%) were mutation carriers (63 MLH1, 51 MSH2). The AUC was 0.95 (95% CI: 0.93–0.97) for MMRpro, 0.87 (95% CI 0.83–0.91) for the Barnetson model, 0.87 (95% CI 0.83–0.91) for PREMM1,2, and 0.75 (95% CI 0.69–0.80) for the Wijnen model (p<0.001). Testing thresholds and specificity at 100% and 90% sensitivity for each model were: 0.001/17% and 0.33/89% for MMRpro, 0.01/9% and 0.07/59% for Barnetson, 0.05/5% and 0.11/58% for PREMM1,2. Sensitivity and specificity of RBG were 86% and 14%, respectively. Calibration was 0.92, 1.05, 0.50, and 1.25 for PREMM1,2,Barnetson, Wijnen, and MMRpro, respectively. Conclusions: In a population of individuals at risk of Lynch syndrome, the MMRpro model has the largest AUC, although the Barnetson and PREMM1,2 model also show adequate discrimination. Any of the models perform better than the RBG and provide quantitative risk estimation of finding a MLH1/MSH2 mutation useful in genetic counselling. No significant financial relationships to disclose.
A single genomic HER2 assay for the protein expression and the DNA amplification status
Authors: Carpentier S. , Bertucci F. , Tagett R. , Finetti P. , Jacquemier J. , Le Doussal J. M. , Hermitte F. 
Published: August 2020

e22082 Background: Currently, up to 20% of HER2 tests in breast cancer may be inaccurate. To address this situation, the 2007 ASCO/CAP guideline for HER2 testing recommended using standardized IHC (for HER2 protein expression) and FISH (for HER2 DNA amplification) in a sequential testing algorithm. However, test standardization and equivocal results still remain an issue potentially leading to inappropriate treatment decisions. A new genomic HER2 (gHER2) assay was designed to address these issues. Methods: Frozen samples of 152 primary breast cancers with a positive or negative IHC result (126 IHC 0+ and 26 IHC 3+ determined by tissue microarray) were profiled using Affymetrix U133 Plus 2.0 gene chips. Recursive feature elimination algorithm by support vector machines (RFE-SVM) was used to select probesets correlated to IHC status; to compute a genomic HER2 (gHER2) expression index; and to set a threshold discriminating between IHC 0+ from IHC 3+ samples. This gHER2 assay was validated in 4 independent datasets (n=309) and tested for its ability to reclassify HER2 equivocal cases (IHC 2+, n=21). The assay was standardized using Ipsogen MapQuant Dx. Results: RFE-SVM identified 6 known and 1 unknown genes correlating with the IHC status, all located within the HER2 DNA amplicon. The gHER2 assay gave unequivoqual HER2 status in 96.2 % of samples. The genomic HER2 assay was concordant with IHC status in 94.5% of cases. 95 % of samples with equivocal IHC status (IHC 2+) could be re-classified in accordance with FISH status in 95 % of cases. Conclusions: We could identify a genomic signature of both HER2 DNA amplification and HER2 protein expression. A new standardized genomic HER2 assay was derived that matches the ASCO/CAP guideline requirements of 95 % concordance with validated techniques. It may contribute to reduce the number of equivocal results. [Table: see text]
Intra tumoral CD8-mediated type 1 anti-tumor responses and differentiated triple negative (TN) and HER-2 advanced breast cancer under neoadjuvant chemotherapy
Authors: Marana H. C. , Benevides L. , Tiezzi D. G. , Andrade J. M. , Brito L. G. , Schiavon V. F. , da Silva J. S. 
Published: August 2020

e22106 Background: To determine the predictive value of CD8 T cells tumor infiltrated lymphocytes (TIL) for treatment response in locally advanced breast cancer (LABC) primarily submitted to neoadjuvant chemotherapy. CD8 T cell-mediated immune responses fall into two distinct types based on effector cell-derived cytokine production. Type I CD8 T cells (Tc1) produce IFN-γ, whereas type 2 cells (Tc2) secrete IL-4, IL-5, IL-10, and GM-CSF. Methods: CD8 T cell, FoxP3 and stimulated anti-CD3 production of IFN-γ were quantitatively measured in the TIL sample from 22 consecutive patients with LABC using a FACS cell analyzing and enzyme immunoassay. Neoadjuvant treatment was given to all patients as Docetaxel 75 mg/m2 and Epirubicin 60 mg/m2 i.v. infusion, for one day, each 21/21 days. Results: Univariate analysis showed that CD8 T cell, FoxP3 and production of IFN-γ are more pronounced in Triple Negative (TN) tumor (39±4.1%, 44,15±11.2% and 17.66±9.18% respectivilly) than HER-2 tumor (30.2±3.4%; 31,7±2.62% and 9.69±6.07%) and this difference was not significantly changed by the neoadjuvant chemotherapy. Conclusions: The results suggest that mammary tumor growth and progression were dependent, in part, on effector cell-derived IFN-γ. Here we postulated that this condition was associated to response to neoadjuvante chemotherapy and was distinct expressed by different gene type of tumor breast. This suggested that CD8-mediated type 1 antitumor responses cannot only promote accumulation of distinct endogenous CD8 T cell subpopulations, but also facilitate and preferentially modulate their localization kinetics, persistence, states of activation/differentiation, and function within the primary tumor environment of tumor response/regression. The TN tumor was more chemo-sensitive and had high IFN-γ production. The action way could be done by CD44 high, FoxP3 and TGFβ1 and the activatioin of Treg in TIL that help in the tumor progression. The chemotherapy breaks this and facilitated the tumor regression. These studies offer insight into potential mechanisms for enhancing T cell-based immunotherapy in breast cancer. [Table: see text]
Is age at the time of surgery a predictor of biochemical failure following radical prostatectomy?
Authors: Badalato G. , Barlow L. , Benson M. , McKiernan J. 
Published: August 2020

e22110 Background: Preoperative PSA level, Gleason score, and tumor stage have all been shown to influence risk of recurrence after radical prostatectomy. Increasing age has been associated with more indolent behavior in some cancers. This study evaluates the effects of age at surgery on recurrence-free survival in prostate cancer patients at a single institution stratified by established preoperative risk factors. Methods: Using the Columbia Urologic Oncology Database, a retrospective analysis of 3,736 men treated with open or robotic-assisted laparoscopic radical prostatectomy for prostate cancer from 1988 to 2008 was conducted. Patients were divided into two groups by age at the time of surgery, and recurrence-free survival rates were analyzed using Kaplan-Meier survival curves. The subgroups were stratified by preoperative PSA level, biopsy Gleason score, and clinical stage; multivariate analyses with cox proportional hazards models were used to further identify independent predictors of recurrence. Recurrence was defined as a single PSA level of 0.2 ng/ml or greater at least 28 days after surgery. Results: 1,984 patients were divided into groups 1 (n=1,325 age 40–64) and 2 (n=659 age ≥65). Five-year recurrence-free survival rates were 80.6%(CI: 78.0–82.9%) and 75.6%(CI: 71.5–79.1%) for groups 1 and 2, respectively. In the univariate model, advanced age was significantly associated with an increased overall risk of recurrence (HR 1.30, p=0.012). However, in multivariate analyses accounting for PSA, Gleason score, and clinical stage, age was not shown to be an independent predictor of recurrence (HR 1.04, p=0.76). In a subset of patients with low-grade cancer (Gleason score 2- 6), advanced age was associated in a univariate analysis with an even greater relative risk of recurrence (HR 1.47, p=0.032). However, this was not significant in the multivariate model (HR 1.27, p=0.21). Conclusions: Older patients who undergo radical prostatectomy for prostate cancer appear to have an increased risk of recurrence, which is most notable in patients with low-grade disease. However, age is not an independent predictor of recurrence when accounting for PSA, grade, and stage. No significant financial relationships to disclose.
MUC16/CA125 specific targeting of critical signaling kinases in ovarian cancer
Authors: Manson R. G. , Thapi D. , Ma X. , Rosales N. , Spriggs D. 
Published: August 2020

e22112 Background: The serum marker CA125 is known to originate from the transmembrane protein, MUC16; however, the biological importance of MUC16 is not known. Our lab previously created fluorescent-labeled transfectants, containing truncated versions of the MUC16 molecule. MUC16 expression vectors were transfected into CA125-negative ovarian cancer cell lines. These cell lines have allowed for a unique means of studying MUC16 biology. In vivo data suggest that MUC16 drives a more aggressive phenotype in ovarian cancer, including increased growth, invasion and dissemination. MUC16 transfection is also associated with the upregulation of specific signaling kinases, including EGFR, Erk, Src and Akt. To validate these findings and explore the potential of MUC16 signal targeting through inhibition of downstream kinases, we are examining the effect of a kinase-directed siRNA library on the A2780 ovarian cancer cell line transfected with a carboxy-terminus MUC16-expression vector. With the identification of kinases important to the survival of CA125+ ovarian cancer tissue, we will identify molecular targets for novel, tumor-specific therapies. Methods: Pilot plates containing candidate kinase-directed, negative control and total kill siRNA (Qiagen) were applied to MUC16+ and MUC16- cells. Cells were grown in culture in parallel 96-well plates. Immunofluorescent readings and Alamar Blue viability staining were performed at 72 hours. Results: Transfection reagents and nonspecific siRNA did not substantially change the survival of fluorescent protein expression in MUC16± cells; while hrGFP and mCherry proteins were decreased by ∼75%. None of the final transfection conditions altered Alamar Blue survival. Preliminary studies suggest that siRNA against AKT2 and SRC had specific inhibition against MUC16+ cells compared to MUC16- controls. Studies of three additional AKT2 siRNAs and a full complement of 714 human kinases are now being tested. Kinases with MUC16-specifc effects will be confirmed with three additional siRNAs. Conclusions: Dual fluorescence isogenic cell lines can be used to explore the kinase profile of MUC16+ ovarian cancer cells. Identified target kinases will enhance our knowledge about MUC16/CA125 biology and provide future leads for the treatment of ovarian cancer. No significant financial relationships to disclose.
p53 as a marker of prognosis in African American (AA) women with breast cancer
Authors: Dookeran K. A. , Dignam J. , Ferrer K. , Sekosan M. , McCaskill-Stevens W. , Gehlert S. 
Published: August 2020

e22119 Background: Prior reports suggest that p53 may be of prognostic value in AA women with breast cancer. However, it remains to be determined whether p53 status would add prognostic value beyond the commonly used factors of stage and Intrinsic Subtype Classification (subtype). We evaluated p53 status as a prognostic factor among AA women treated at an urban community hospital. Methods: Cox proportional hazards regression models [results reported as hazard ratios (HR) with 95% confidence intervals (CI)] were used to select and evaluate prognostic factors [including stage, age, tumor grade of differentiation (grade), p53 status, subtype, & ER/PR status] for all-cause mortality in 331 consecutively treated AA women with breast cancer [42 months follow-up] and known subtype [luminal A = ER+, &/or PR+, & HER2-; luminal B = ER+, &/or PR+, & HER2+; HER2+ = ER-, PR-, & HER2+; basal = ER-, PR-, HER2-, cytokeratin (CK) 5/6+ &/or HER1+; & unclassified = negative for all 5 markers] and p53 [Pab1801 antibody] immunohistochemical status. Results: Tumors in 28% of women were p53+ and there were no chemotherapy and radiation treatment differences according to p53 status. However, 59% of p53+ women were ER/PR negative [Odds Ratio (OR), 0.37; 95% CI, 0.22–0.54; p=0.0003] and hence endocrine therapy was significantly less frequent in p53+ women [OR, 0.40; 95% CI, 0.23–0.69; p=0.0008]. p53+ tumors were also significantly more likely to be grade 3 [OR, 4.35; CI, 1.33–14.14; p=0.013]. Baseline prognostic factors were: stage [(II-IV/I) HR, 2.29; 95% CI, 1.86–2.81; p<0.0001]; age [HR, 1.003 per year; 95% CI, 0.99–1.02; p=0.697]; grade [(high/low) HR, 1.70; 95% CI, 1.22–2.37; p=0.0008]; p53 status [(±) HR, 1.76; 95% CI, 1.15–2.72; p=0.012]; subtype [(all other/luminal A) HR, 1.33; 95% CI, 1.14–1.55; p=0.0004]; ER/PR status [(±) HR, 0.47; 95% CI, 0.32–0.69; p=0.0001]. Cox multivariable models indicated that p53 status [HR, 1.59; 95% CI, 1.01–2.51; p=0.044] remained a significant prognostic factor when considered with stage [HR, 2.20; 95% CI, 1.71–2.84; p<0.001] and subtype [HR, 1.24; 95% CI, 1.04–1.49; p=0.016] and the other above-mentioned factors. Conclusions: Study results indicate that p53 status should be included with stage and subtype as markers to assess prognosis in AA women with breast cancer. No significant financial relationships to disclose.
HMPS (2-hydroxy-4-methoxyphenylstilbene), a stilbene derivative of rhapontigenin, and cell death by mitochondrial apoptotic pathway in breast cancer cells
Authors: Jung J. , Park H. , Jung H. , Eun Y. , Kim J. , Chae Y. , Yang J. , Lee Y. , Aiyar S. E. , Santen R. J. , Kim S. 
Published: August 2020

e22130 Background: Breast cancer with resistance to clinical therapy is a significant threat to live of recurrent breast cancer patients, and chemo-resistant breast cancer is increasing rapidly. During last several decades, natural stilbenoids have been studied on anticancer effects in vitro and in vivo, and resveratrol is the most famous stilbene as a leading compound in the studies of anticancer compounds derived from plants. HMPS (2-hydroxy-4-methoxyphenylstilbene) is an analogue derived from rhapontigenin (3,5,3'-trihydroxy-4'-methoxy-trans-stilbene), which is a stilbene of herbal plant Rheum undulatum. TMS (2,3',4,5'-tetramethoxystilbene), an another stilbene analogue from rhapontigenin, was reported potent anticancer effect on tamoxifen-resistant MCF-7 cells. In this study we investigated inhibitory effect of HMPS on proliferation of breast cancer and a potential for a new therapeutic candidate. Methods: We examined cell viability of MCF-7 and MDA-MB-231 by MTT assay after exposure to various concentrations of HMPS. Apoptotic cell death induced by HMPS was investigated by florescence microscopy, cell cycle analysis and western blotting. Results: Cell viability of breast cancer cells after 24 h exposure to HMPS decreased significantly, and both ER-positive and ER-negative breast cancer cells responded to HMPS. HMPS induced nucleus fragmentation and G2/M arrest followed by sub-G1 accumulation of apoptotic cells in time- and dose-dependent manner. During the process of cell death induced by HMPS, mitochondrial membrane potential was disturbed and caspase-3 and PARP cleavage were observed. Moreover, HMPS decreased cell number of LTED MCF-7 cells(Long term estradiol deprived cell) effectively. Conclusions: Our results demonstrates that proliferation inhibitory effect of HMPS is about 50-fold more potent than those of rhapontigenin and furthermore HMPS also inhibits cell growth of LTED cells which are difficult to treat therapeutic agents. Therefore, HMPS may be a potential therapeutic candidate to treat the recurrent breast cancer by alone or combination with other conventional anticancer agents. No significant financial relationships to disclose.