bluuebunny/crossref_metadata_2025_split huggingface dataset:
Heterogeneity of HDAC3 expression and its modulation by valproic acid in glioblastoma cells.
Authors: Engelhard Herbert H. , Lu Yi , Rayfield Corbin , Yue Lili , Lakka Sajani , Mahmud Nadim 
Published: March 2020

e13038 Background: Glioblastoma (gbm) is a devastating malignancy and a therapeutic challenge. Our group and others have observed long-term gbm survivors treated with valproic acid (VPA), a histone deacetylase (HDAC) inhibitor. While in vitro studies of HDAC inhibitors have been promising, clinical trials and retrospective analyses have been disappointing. We hypothesize that HDAC3 may be a more specific target or treatment marker for gbm. Since limited data is available pertaining to HDAC3 and its modulation in gbm, our goals were to: 1) develop methods for determining HDAC3 expression and localization in gbm cells, 2) study HDAC3 heterogeneity among different gbm cell lines, including one in low passage, and 3) test the effect of VPA treatment on HDAC3 expression of gbm cells in vitro. Methods: Three established gbm cell lines (U-87, U-251 and E297), were cultured using standard technique. HeLa cells were used as the positive HDAC3 control. Cells were stained using 2 different antibodies, and analyzed using immunofluoresence microscopy (IFM), and Western blotting (with β-actin standards). IgG was used for negative isotype controls. Cell counts, viability, and H&E staining for changes in morphology were also recorded. Lysates for Western blots were made from whole cells, nuclei, and cytoplasmic preparations. Band quantification was performed using the ImageJ software. Cells in exponential growth were treated for 48 hours with 10 mM VPA, and compared to untreated cells. Results: Untreated cells studied for HDAC3 using IFM showed nuclear and some cytoplasmic staining in all 4 lines, which varied according to cell type and morphology. Western blot analysis confirmed the variability of HDAC3 expression seen by IFM. The effect of VPA treatment on gbm HDAC3 expression (cytoplasmic and nuclear) was seen to be highly variable, ranging from marked down-modulation in some gbm cells, to minimal effect in others. Conclusions: These data provide the groundwork for studying HDAC3 levels in patients undergoing therapy with HDAC inhibitors. The effect of VPA on the HDAC3 expression of gbm cells is heterogeneous; individualized assessment of such effect will be necessary in delineating any subset of patients that may respond to VPA treatment.
ChemoID assay for anaplastic spinal ependymoma.
Authors: Valluri Jagan , Claudio Pier Paolo , Mathis Sarah E , Alberico Anthony , Kimmey Gerrit A. , Mogul Mark Jeffrey , Sehgal Rajesh , Chowdhary Aneel A. , Tirona Maria R. B. Tria , Dougherty Thomas 
Published: March 2020

e13040 Background: Cancer stem-like cells (CSLCs) can resist certain chemotherapies, thereby causing relapse of the disease. Thus, development of a test that identifies the most effective chemotherapy management offers great promise for individualized anticancer treatments. Methods: We have developed ChemoID, an in vitro chemotherapy assay, designed to predict the sensitivity and resistance of a given patient’s CSLCs and bulk of tumor cells to a variety of chemotherapy agents. Results: A 5-month female (Patient A), and a 21-year old male (Patient B), both affected by anaplastic WHO grade 3 ependymoma were screened using the ChemoID assay. A range of concentrations flanking the clinical doses for vincristine, carboplatin, cyclophosphamide, etoposide, cisplatin, methotrexate, cytosine arabinoside, oxaliplatin, irinotecan, avastin, procarbazine, lomustine, and busulfan were tested. The CSLCs and bulk of tumor ependymoma cells of Patient A were found moderately sensitive (30% cell kill) to only cisplatin and procarbazine, both as single agents and in combination. Following 6 cycles of vincristine, carboplatin, cyclophosphamide, etoposide, and cisplatin in various combinations, Patient A rapidly progressed and proton beam therapy was recommended. The CSLCs and bulk of tumor ependymoma cells of Patient B were found highly sensitive (80% cell kill) to the combination of irinotecan and avastin, which resulted in 18 months free of disease progression. After recurrence, chemotherapies were tested in combination with benzyl-isothiocyanate (BITC), a bioactive natural food component contained in extracts from cruciferous vegetables. BITC greatly increased the chemosensitivity in Patient B to the combination of irinotecan and avastin, resulting in average 90% CSLCs and bulk of tumor ependymoma cell eradication. MRI scan of Patient B showed over 50% tumor regression after two months of treatment with a combination of irinotecan, avastin, and supplements containing BITC. Conclusions: Our findings suggest that no patients are alike in their chemotherapy response. ChemoID assay that measures cell death of CSLCs and bulk of tumor cells from patient biopsies to standard chemotherapy agents in vitro is a feasible approach to personalized anticancer treatments.
Kinin system and α-2-macroglobulin of liquor in treatment of gliomas by local chemotherapy.
Authors: Kozlova Larisa , Frantsiyants Yelena , Gorbunova Tatiana 
Published: March 2020

e13042 Background: Local administration of cytostatic into the bed of removed tumor during operation will require a smaller quantity of the drug, which will reduce its toxicity and ensure entire use of cytostatic according to its purpose, without any essential impact on other organs. It is relevant to study the kallikrein-kinin system (KKS) and α-2-macroglobulin (α-2M) of liquor after intraoperation local chemotherapy (ILCT) of gliomas in dynamics. Methods: Liquor of 40 patients (26 men and 14 women, 39-60 years old, T2-3NõMo) who received ILCT was studied. ILCT consisted in 0,2 mg carboplatin being placed in a container of hemostatic sponge, with hemostasis performed next, and the former put into the bed of the tumor removed. The results were compared to the check liquor (CL) of 8 patients, for whom malign and inflammatory processes in brain has been ruled out and whose age and sex characteristics are comparable to the oncological patients. Results: Before the treatment (the background), kallikrein (K) and prekallikrein (PK) content was higher than that in CL in 38.3 and 2.6 times, respectively. The activity of kininase 1 (K1) was 0.989±0,1 mcM/ml (it was not found in the CL). The activity of α-2M is increased 9 times as compared to the CL. On day 1 after ILCT, PK/K increased in 4.4 times, K/K1 and K/α-2M went down by 1.6 and 1.5 times. On day 7, PK/K decreased in 1.7 times, K/α-2M – in 2 times, while K/K1 remained at the same level. On day 14, the proportion of PK/K was maintained, and K/K1 and K/α-2M increased in 3.4 and 2.5 times. After 1.5 months, PK/K did not change, while K/K1 and K/α-2M reduced by 1.9 times each. The results demonstrate high reactivity of liquor against ILCT and activation of restorative processes within days 1-7 after treatment. The imbalance observed on day 14 was improved in 1.5 months after ILCT. Conclusions: ILCT fosters execution of protective function of proteins under study within not less than 1.5 months after treatment. Studying the KKS and α-2M in liquor has prospects for controlling the therapy administered.
Textiloma mimicking recurrent glioblastoma: A case report.
Authors: Garciarena Pedro , Anderson Mark Daniel , Hamilton Jackson , Schellingerhout Dawid , Fuller Gregory N , Sawaya Raymond , Gilbert Mark R. 
Published: March 2020

e13044 Background: Although uncommon, materials used for dissection or to achieve hemostasis during neurosurgical procedures can cause an inflammatory reaction called textiloma that can mimic recurrent tumor. We report a case where advanced magnetic resonance (MR) imaging suggested tumor, further mimicking previous reports of clinical and conventional MR patterns of tumor progression. Methods: A 61 year-old woman underwent an initial resection of a temporal lobe glioblastoma, followed by conventional chemoradiation, although maintenance temozolomide was stopped secondary to myelotoxicity. A nodular enhancing mass near to the surgical cavity was noted 10 months after diagnosis, leading to further evaluation with advanced MR imaging. Results: Brain MRI with advanced imaging studies including MR Spectroscopy (MRS), and perfusion imaging by dynamic contract-enhanced (DCE), and dynamic contrast susceptibility (DSC) were performed. There was progressive enlargement of an enhancing nodule in the wall of the resection cavity. MRS demonstrated an elevated choline-to-creatine ratio (approximately 2:1), while the DSC imaging demonstrated a mild amount of leak with a corrected cerebral blood volume value of about 1.93, and negative enhancement integral value 1.67. DCE demonstrated mild vascularity and mild-to-moderate leak, with an elevated peak enhancement integral value of ~22%. These imaging findings suggested recurrent GBM rather than post-treatment change. The mass was resected. Pathologic review showed both textiloma and radiation necrosis. Recurrent glioblastoma was not identified. Conclusions: Although very unusual, textiloma should be considered in the differential diagnosis of any growing mass lesion close to a surgical bed. Reparative granulomata show many of the hallmarks of cancer, including neovascularization, rapid growth and high lipid turnover, making it difficult to distinguish such lesions from true neoplasm, even with advanced imaging, without resorting to histology. Advanced imaging performs best in distinguishing conventional post-treatment change from tumor recurrence. This case highlights the major influence of surgical technique on postoperative imaging appearance.
Clinical, pathologic, and radiologic response in patients with neurofibromatosis type 2 (NF2) undergoing bevacizumab treatment.
Authors: Coleman Niamh , Greally Megan , O' Riordan Lynda , Doyle Teresa , Breathnach Oscar S. , Grogan William 
Published: March 2020

e13049 Background: NF2 is a rare inherited genetic condition characterized by multiple "benign" tumors in the peripheral and central nervous system. Profound hearing loss as a result of bilateral vestibular schwannomas is a major debilitating complication with a profound effect on quality of life. Vascular endothelial growth factor (VEGF) has been shown to be produced by vestibular schwannoma tumor cells. Bevacizumab, a humanized monoclonal antibody against VEGF, has been proposed as a treatment for patients with advanced NF2, who are poor candidates for surgery and radiation therapy. The aim of this study was to investigate and assess the benefit of treatment with bevacizumab in the treatment of patients with advanced NF2. Methods: Five patients with inoperable NF2 were assessed prospectively from 03/12-01/13. Pathological assessment of tissue for vascularity was performed in each of the patients’ tissue. Objective assessments, including audiogram and MRI brain, were performed pre- and during therapy. Qualitative assessments were undertaken to assess subjective response to treatment. Results: We identified 5 patients with advanced NF2 [n=5], 3 female 2 male with a median age 40 years [range 28 – 52] as outlined in the Table. Tumor-volume reduction of >20% was noted on MRI imaging in the vestibular schwannomas of 2 patients. 2 patients showed objective improvement on audiogram with subjective hearing improvement also noted. Other subjective responses noted include increased energy levels, less headaches, better concentration. Conclusions: Bevacizumab treatment in patients with advanced NF2 resulted in some symptomatic improvement, tumour shrinkage, improvement in disability. Patients with vestibular schwannomas appeared to receive the most benefit, and so in terms of palliation, the benefits may be most significant in patients with specific disabilities, such as hearing loss. [Table: see text]
Development of a new equation to estimate GFR in cancer patients.
Authors: Chu Michael Patvin , McCaw Larissa , Stretch Cynthia , Hanson John , Kuzma Michelle , Damaraju Vijaya L. , Baracos Vickie E. , Sawyer Michael B. 
Published: March 2020

e13503 Background: Renal function affects chemotherapy pharmacokinetics. Carboplatin dosing by Calvert’s formula is more pharmacologically rational, but requires an accurate glomerular filtration rate (GFR). Calvert argues that this requires measuring GFR (mGFR) instead of an estimated GFR (eGFR). Considering skeletal muscle is the major source for creatinine, this study looks to develop a new eGFR equation in cancer patients using lean body mass (LBM). Methods: We prospectively followed 22 stage IV cancer patients (10 female, 12 male; median age 69) who received carboplatin. mGFR by 24 hr creatinine clearance was compared to eGFR by Wright, Cockcroft-Gault (CG), CKD-EPI, MDRD and CT-determined LBM (eGFR = [Muscle Surface Area X 42]/CR). Simulated carboplatin dosing with each eGFR was then compared retrospectively in 100 Non-Small Cell Lung Cancer (NSCLC) patients for accuracy. Results: MDRD, CG, and Wright equations correlated variably with mGFR (R2 0.47, 0.57, and 0.69 respectively). Conversely, mGFR strongly correlated with LBM eGFR (R2 0.84). The Table compares eGFR calculations with mean residual error. In simulated carboplatin dosing of 100 stage IV NSCLC patients using LBM and CG eGFR, the mean residual error of the CG-determined carboplatin dose was 10% (0.5% min, max 39.7%, median 9.3%), assuming the LBM eGFR was better at estimating eGFR. This means that in approximately half of patients, carboplatin dose may be incorrect by CG if the new LBM eGFR method is truly more accurate. Conclusions: We propose a new formula for eGFR in cancer patients that appears superior to current formulas and may have implications for chemotherapy efficacy and toxicity. Studies to validate this formula are under way. [Table: see text]
Co-targeting PI3K and ras pathways in trastuzumab resistance.
Authors: Gray Mike J , YangKolodji Gloria Wangrong , Tripathy Debu 
Published: March 2020

e13515 Background: HER2+ breast cancers (BC) account for 20–25% of invasive BC and are associated with an aggressive phenotype and poor patient outcome. The development of trastuzumab and other HER2-targeted therapy dramatically improved outcomes for HER2-positive BC, but most patients with advanced HER2+ BC will eventually become resistant to treatment, underlying the importance of developing alternative or combination treatments. Alterations in the PI3K/mTOR/Akt pathways are cited as contributors to the development of trastuzumab resistance, however targeting these kinases as single agents has yielded less than expected clinical results. This suggests that combinational treatment with other kinase pathway inhibitors may be required, including those targeting the Ras/MAPK pathway, which is typically not mutationally activated in breast cancer. Methods: Trastuzumab resistant HER2+ BC cell lines were subjected to dose responses with the mTOR inhibitor everolimus or the AKT inhibitor MK-2206 alone or in combination with the MEK 1/2 inhibitor GSK212 and changes in EC50s determined by MTT assay. Western blot analysis was performed to assess changes in the mTOR/AKT/ MAPK pathways or apoptotic regulators accompanying single or combination treatments. Results: In 4 of 5 trastuzumab-resistant HER2+ BC cell lines, each lacking activating mutations in the Ras/Raf pathway, combination treatment with everolimus or MK-2206 with GSK212 had significantly greater efficacy than by either inhibitor alone. Furthermore, combinational treatment targeting the mTOR and AKT pathways showed only an additive effect, and was much less effective than targeting both the MEK and PI3K pathways. Western analysis showed that AKT and mTOR inhibition caused a transient increase in ERK 1/2 activity in sensitive cell lines, suggesting that treatment by mTOR or AKT inhibitors activated critical survival pathways in the MAPK signaling that were blocked by GSK212. Conclusions: Treatment with mTOR or AKT inhibitors in combination with MEK inhibitors can act in a synergistic manner with greater efficacy than each inhibitor alone. Moreover, ERK 1/2 activity may serve as a predictive biomarker in trastuzumab-refractory patients treated with mTOR/MEK or AKT/MEK doublet therapy.
Efficacy of the phosphoinositol 3 kinase (PI3K) inhibitor GDC-0941 in patient- and cell-line-derived xenografts of dedifferentiated liposarcoma (DDLPS).
Authors: Li Haifu , Wozniak Agnieszka , Van Den Bossche Karel , Van Looy Thomas , Wellens Jasmien , Stas Marguerite , Floris Giuseppe , Friedman Lori , Debiec-Rychter Maria , Sciot Raf , Schoffski Patrick 
Published: March 2020

e13528 Background: Analysis of primary clinical specimens of DDLPS revealed evidence of AKT activation in 27% of cases (Gutierrez A, et al. Aberrant AKT activation drives well-differentiated liposarcoma. PNAS 2011; 108(39):16386-91). We evaluated the efficacy of the PI3K inhibitor GDC-0941 (GDC) alone and in combination with doxorubicin (DOX) in patient- and cell-line derived DDLPS xenografts with proven PI3K/AKT pathway activation. Methods: NMRI nu/nu mice were either injected bilaterally with 1x107SW872 cells or transplanted with human DDLPS (UZLX-STS3). Animals were randomized to 4 groups (6 mice/group) and treated for two weeks: vehicle, DOX (i.p., 1.2mg/kg/biw), GDC (p.o., 75mg/kg/qd), GDC+DOX (same dose/schedule as single agents). Efficacy was assessed by tumor volume assessment, Western blotting and histological evaluation (H&E). Mitotic and apoptotic effects were confirmed using immunostainings for phospho-histone H3 (p-H3) (for proliferation), and cleaved caspase 3 (CC3) (for apoptosis). Results: GDC and GDC+DOX significantly delayed the growth of both xenografts (74% decrease under GDC and 77% under GDC+DOX in SW872, 67% under GDC and 56% under GDC+DOX in UZLX-STS3, vs. control). GDC and GDC+DOX treatment resulted in reduced mitotic activity and increase in apoptosis, as compared to untreated tumors in both models (Table). The activation of PI3K signaling was nearly completely suppressed in the GDC and GDC+DOX treated groups in SW872, whilea weak activation of the pathway was still observed in UZLX-STS3. Conclusions: GDC-0941 has anti-tumor activity, decreases tumor proliferation and induces apoptosis in DDLPS with PI3K/AKT activation. The GDC+DOX combination did not show additive effect in vivo. [Table: see text]
Synergy with combination of AKT inhibitor (MK-2206) and paclitaxel in head and neck squamous cell carcinoma.
Authors: Ahmed Omar , Kuo Wen-Liang , Nagilla Madhavi , Messer Jeannette S. , Boone David L. , Sharifi Marina , Macleod Kay , Cohen Ezra E.W. 
Published: March 2020

e13532 Background: Though there have been many advances in the treatment of HNSCC, the 5-year survival rate remains at 50%. This is partly due to molecular alterations such as the activation of the PI3k-AKT pathway in HNSCC, which has been associated with treatment failure. We stained human tissue samples and saw increased expression of p-AKT and pS6 in HNSCC compared to normal and dysplasia. This led us to investigate the role of the AKT inhibitor MK-2206 in combination with an already established chemotherapeutic agent, paclitaxel. Methods: Cell viability and Combination index were used to determine synergy in 4 HNSCC cell lines. PI exclusion and Annexin/PI staining were used to determine cell death and apoptosis, respectively. Autophagy was measured through changes in LC3BII protein levels using western blot analysis and a stably transfected GFP-LC3BII cell line was used to detect changes in protein localization under confocal microscopy. Results: All cell lines were sensitive to paclitaxel while only some were sensitive to MK-2206. However, there was effective synergy to the combination treatment in all cell lines. Apoptotic synergy was observed in some cell lines indicating that the combination stimulates cell death in the appropriate context. In one autophagy competent cell line, MK-2206 induced robust autophagy while paclitaxel blocked a latter step in the pathway leading to an accumulation of autophagosomes prior to induction of apoptosis. Conclusions: Paclitaxel and MK-2206 appears to be an effective combination in HNSCC possibly through their effect on autophagy trafficking, leading to an accumulation of autophagosomes and eventual apoptosis. Clinical trials examining this combination are warranted.
Heat-shock (H-S) and trabectedin efficacy in human soft-tissue sarcoma (STS) cells in vitro.
Authors: Kampmann Eric , Harnicek Dominique , Cardoso Martins Ana Sofia , Eppink Berina , Gallmeier Eike , Lindner Lars , Kanaar Roland , Issels Rolf D. 
Published: March 2020

e13540 Background: Regional hyperthermia improves response and survival when combined with chemotherapy in patients with high-risk STSs (Issels, R.D. Lancet Oncol 2010). Trabectedin is the first marine-derived antineoplastic drug approved in Europe for the treatment of advanced STS after failure of anthracyclines and ifosfamide, or for patients who are unsuited to receive these drugs. Trabectedin’s cytotoxicity is associated with the induction of lethal DNA double-strand breaks (DSB). The rationale to combine trabectedin with H-S is that heat-exposure sensitizes tumor cells by inhibiting the repair of induced DSBs (Krawczyk, P.M. PNAS 2011). Methods: Combinations of trabectedin and H-S at clinically relevant temperatures were examined in 3 different human cell lines: Osteosarcoma (U2Os), liposarcoma (SW872) and synovial sarcoma (SW982). Cells were treated with trabectedin at the dose of 500-4000 pM for 3 hours. H-S was applied in an incubator at 41.8°C and 43°C for 90 or 150 min before, during or after trabectedin incubation. Cytotoxicity was assessed measuring clonogenic survival of cells. Expression of BRCA2, which recruits homologous recombination repair recombinase Rad 51 to DSBs, was measured by Western Blot (WB). Recruitment of Rad 51 and the amount of gH2AX positive DSB-repair-foci were analysed by immunocytochemistry (ICC). Results: All cell lines showed reduced viability after increasing doses of trabectedin at 37°C. Combined treatment with trabectedin and H-S additionally enhanced cytotoxicity of trabectedin with strongest effects observed after simultaneous administration of both. WB-analysis showed strong heat-dependent reduction of BRCA2 expression. ICC revealed that recruitment of Rad 51 to DSBs was reduced after heat exposure at 41.8°C and abolished after exposure at 43°C. Accordingly, combined treatment significantly increased the amount of cells with severe DNA damage (>50 DSBs). Conclusions: Combined treatment with trabectedin and H-S in vitro resulted in significantly enhanced cytotoxicity that was accompanied by elevated DNA-damage in term of DSB-accumulation. The mechanisms of interaction between trabectedin and H-S concerning DNA repair are under investigation.