bluuebunny/crossref_metadata_2025 huggingface dataset:
Patient selection for anti-EGFViii therapies in glioblastoma multiforme (GBM): Use of circulating tumor DNA.
Authors: Bahassi El Mustapha , Salkeni Mohamad Adham , McPherson Christopher , Warnick Ronald , Rixe Olivier
Published: March 2020

e13023

Background: Several strategies have been recently developed to target EGFRvIII in Glioblastoma Multiforme (GBM), including vaccines (CDX-110) and antibody-drug conjugates (AMG595) and represents a new challenging therapeutic avenue with potential great clinical benefits. EGFRvIII is a truncated extracellular mutant of the EGF receptor (EGFR) found in about a third of GBMs, confers enhanced tumorigenic behavior and is associated with chemo- and radio-resistance. GBM patients testing positive for EGFRvIII have a bleaker prognosis than those who do not. Methods: We developed a strategy to detect EGFRvIII deletion in the circulating tumor DNA. The purpose of this study is to identify a simple and robust biomarker from the peripheral of patients diagnosed with GBM in order to screen patients for the EGFRvIII deletion. 11 patients have been included in this study. Results: The circulating DNA status for EGFvIII correlates with the analysis performed on the respective tumors samples, and its level seems to be correlated with the extend of the tumor resection. Conclusions: This semi-quantitative blood biomarker may represents a strategy to 1) screen patients for anti-EGFRvIII therapy 2) monitor the therapeutic response to specific targeted therapies.
Risk factors for venous thromboembolism in elderly patients with glioblastoma.
Authors: Hashemi-Sadraei Neda , Sharif Roozbeh , Machicado Jorge , Dahiya Saurabh , Ahluwalia Manmeet Singh
Published: March 2020

e13031

Background: Venous thromboembolism (VTE) is a common complication in glioblastoma (GBM) patients. Physicians are reluctant to administer anticoagulants to GBM patients with VTE, particularly to the elderly patients. Methods: After obtaining IRB approval, the Cleveland Clinic Brain Tumor and Neuro-Oncology Center’s database was used to identify patients with histologically confirmed GBM who were diagnosed with VTE and were ≥65 years of age at the time of diagnosis of GBM. Bivariate comparison of demographic and clinical features was performed by occurrence of DVT, using t-test for continuous data and fishers exact test for categorical variables. Multivariate proportional hazard model was developed adjusting for number of comorbidities (none, 1, 2-3, >3), Karnofsky Performance Status (KPS) at presentation (>70, ≤70), age (>75, ≤75), race, treatment (chemotherapy, radiation therapy, tumor resection). All p-values were 2-sided and were considered significant when <0.005. Results: Chart records of 517 patients diagnosed between 1990 and 2010 were included for analysis. Among 517 cases, 99 patients (54% male, median age at presentation 72 years, range 65-92 years) developed VTE. Median follow-up was 6.58 ± 9.58 months. Eighty-nine (90.8%) cases were diagnosed within 3 months of craniotomy. On univariable analysis, risk factors for VTE included lower KPS (HR 0.98, CI 0.96– 0.99, p = 0.001), thalamic tumors (HR 3.11, CI 1.14– 8.50, p = 0.027), and lower extremity paresis (HR 2.24, CI 1.46-3.46, p < 0.0001). Caucasian patients had a lower trend for developing VTE, however this was not statistically significant. On multivariable analysis, lower KPS was found to be related with higher incidence of VTE (HR 0.35, CI 0.18 – 0.66, p = 0.001). Conclusions: Lower KPS is associated with higher incidence of VTE. Further studies are needed to determine risk stratification and whether the use of medical prophylaxis after neurosurgical intervention can lead to decrease in the rates of VTE in elderly GBM patients.
Glioblastoma multiforme metastasis outside the central nervous system: Three case reports and possible mechanisms of escape.
Authors: Boelke Edwin , Matuschek Christiane , Ginsberg Lawrence E. , Prabhu Sujit S. , Budach Wilfried , Hayman Anne , Hamilton Jackson
Published: March 2020

e13035

Background: Primary brain and central nervous system (CNS) tumor incidence is approximately 19 per 100,000 individuals per year in the United States (US) compared with 7 per 100,000 individuals worldwide. The most common intra-axial tumor is gliomas, which account for 32% of all primary CNS tumors and 80% of all malignant tumors of the CNS.The most common diffuse glioma is grade 4 astrocytoma (glioblastoma, GBM), which makes up 54% of diffuse glial tumors. GBM is also the most aggressive brain tumor with poor prognosis.GBM metastases outside the CNS are rare, so therapeutic experience with these types of tumors is limited. Methods: Herein, we present 3 GBM patients with extra-CNS metastasis. Results: One patient developed GBM metastasis in the lung and pleura 5 years after his GBM diagnosis had been confirmed. Another patient who underwent resection of the primary GBM developed disease that extended through the sphenoid to involve the orbit and skull and subsequently invaded the parotid gland and neck nodes 1 year after diagnosis. A third patient developed GBM metastasis in the skull and L5 vertebra 2 years after her primary brain tumor had been resected. Conclusions: The exact mechanism of GBM metastasis outside the central nervous system is not well understood but likely involves the invasion of structures such as bone, lymphatics, and vasculature, especially veins. Above-average survival time and repeated surgical intervention may place GBM patients at higher risk for these unusual metastases.
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.
Lipegfilgrastim: A long-acting, once-per-cycle, glycopegylated recombinant human filgrastim.
Authors: Scheckermann Christian , Schmidt Karsten , Abdolzade-Bavil Afsaneh , Allgaier Hermann , Mueller Udo W. , Shen Wenyan David , Liu Patrick
Published: March 2020

e13548

Background: Lipegfilgrastim is a once-per-cycle fixed-dose glycoPEGylated granulocyte-colony stimulating factor (G-CSF) under development for the prevention of severe neutropenia in cancer patients receiving chemotherapy (CTx). PEGylation of a molecule extends its half-life in the body, requiring less frequent dosing and allowing for administration of G-CSF once per CTx cycle, making treatment potentially less expensive and enhancing patient compliance and safety. Briefly, traditional PEGylation methods use chemical conjugation through reactive groups on amino acids, which may reduce protein activity and result in non-uniform chemical and pharmaceutical properties. Here, we describe a highly site-specific GlycoPEGylation technology for site-directed PEGylation and summarize preclinical findings compared with pegfilgrastim (Neulasta). Methods: Glycosylation sequon scanning was used to identify the glycoPEGylation site with least impact on protein activity. E. coli-expressed G-CSF was selectively glycosylated at natural O-glycosylation sites and a polyethylene glycol (PEG) sialic acid derivative attached using a sialyltransferase (glycoPEGylation technology). Ligand binding affinity was assessed using the BIACORE 3000 system. Biologic activity of lipegfilgrastim was assessed in an NFS-60 cell line proliferation assay vs filgrastim and pegfilgrastim. PK and PD properties were studied in healthy and neutropenic animal models. Results: GlycoPEGylation produces long-acting G-CSF with improved PK profiles. In vitro, lipegfilgrastim had binding affinity and specific activity comparable to pegfilgrastim, and both were lower than non-PEGylated filgrastim. Comparable increases in leukocytes, neutrophilic granulocytes, and monocytes were seen with lipegfilgrastim and pegfilgrastim in rats and monkeys and were consistent with the effects expected for a long-lasting G-CSF. Conclusions: GlycoPEGylation technology platform is used to produce lipegfilgrastim – a novel, biologically active G-CSF with greater structural homogeneity and comparable pharmacologic properties to conventionally PEGylated G-CSFs.
Balugrastim: A long-acting, once-per-cycle, recombinant human albumin-fusion filgrastim.
Authors: Pukac Laurie , Barash Steven , Avisar Noa , Allgaier Hermann , Bock Jason , Mueller Udo W. , Shen Wenyan David
Published: March 2020

e13551

Background: Balugrastim is a once-per-cycle fixed-dose genetic fusion protein composed of human serum albumin (HSA) and granulocyte colony-stimulating factor (G-CSF) in development for prevention of severe neutropenia in cancer patients receiving chemotherapy. Albumin fusion is a clinically validated technology that extends product half-life, allowing for infrequent dosing, better tolerability, lower cost, and improves drug design, potentially lowering immunogenicity risk. Here, we describe the technology used to produce balugrastim and summarize preclinical findings compared with pegfilgrastim (Neulasta). Methods: Design and production of balugrastim was described previously (Halpern et al. Pharmaceut Res 2002;19:1720−1729). Biologic activity of balugrastim was assessed in an NFS-60 cell line proliferation assay vs filgrastim and pegfilgrastim. PK and PD properties were studied in healthy and neutropenic animal models. Results: Albumin fusion produces a long-acting G-CSF with comparable pharmacologic properties to pegfilgrastim. In vitro, balugrastim had binding affinity and cell proliferation activity comparable to pegfilgrastim, and both were lower than non-PEGylated filgrastim on a molar basis. Overall increases in leukocytes, neutrophilic granulocytes, and monocytes were dose dependent and consistent with the effects expected for a long-acting G-CSF with some variation based on the specific animal model used. A single balugrastim dose in BDF1 mice elicited a dose-dependent increase in peripheral granulocytes and mobilized hematopoietic progenitor cells. In cynomolgus monkeys, balugrastim caused an increase in peripheral neutrophils similar to pegfilgrastim, with higher responses after 2nd, 3rd, and 4th doses. In mice, balugrastim had shorter terminal half-life and mean residence time, and faster clearance than pegfilgrastim. In monkeys, terminal half-life of balugrastim was slightly longer than pegfilgrastim. Conclusions: An albumin fusion technology platform was used to produce balugrastim – a novel, biologically active albumin G-CSF fusion protein with greater structural homogeneity and comparable pharmacologic properties to conventionally PEGylated G-CSFs.
Clinical safety of tbo-filgrastim in chemotherapy-induced neutropenia: Integrated analysis of the phase III studies.
Authors: Engert Andreas , Buchner Anton , Mueller Udo W. , Bias Peter
Published: March 2020

e13552

Background: Tbo-filgrastim is a granulocyte colony-stimulating factor (G-CSF) approved for use in the US as a biologic under BLA 351(a) for reducing duration of severe neutropenia in patients with non-myeloid malignancies receiving myelosuppressive chemotherapy (CTx) associated with a clinically significant incidence of febrile neutropenia. Here, we report an integrated analysis of clinical safety data from three Phase III studies. Methods: Safety results from three Phase III studies in breast cancer (placebo-controlled), non-small cell lung cancer, and non-Hodgkin lymphoma were integrated. All safety variables were analyzed using descriptive statistics. The incidence of treatment-emergent adverse events (TEAEs) was compared between treatment groups using Fisher's exact test (2-sided P-values). Changes in laboratory parameters from baseline were compared between the tbo-filgrastim and filgrastim treatment groups using the Wilcoxon test. Results: In the integrated analysis of CTx Cycle 1, 80.2% of 677 patients experienced a TEAE; 16.7% of these were related to study drug. The most commonly reported TEAEs were nausea, alopecia, neutropenia, diarrhea, asthenia, and vomiting, and most due to concomitant CTx. The most commonly reported drug-related TEAEs were bone pain (3.4%), diarrhea (2.2%), asthenia (2.2%), myalgia (1.9%), arthralgia (1.5%), headache (1.2%), and pyrexia (1.0%), all of which are characteristic of G-CSFs. Five patients had serious drug-related TEAEs; none caused reductions in the dose of study drug or scheduled CTx. There were 11 deaths across studies; none was considered drug related. There were no clinically significant differences between tbo-filgrastim and filgrastim in the overall incidence of deaths, serious or severe TEAEs, discontinuations due to TEAEs, or in laboratory safety variables. In all cycles of CTx, the most commonly reported TEAEs in patients treated with tbo-filgrastim correlated with those of the reference filgrastim. Conclusions: Tbo-filgrastim is safe and well tolerated in breast cancer, non-small cell lung cancer, and non-Hodgkin lymphoma patients undergoing CTx, with a safety profile similar to that of the reference filgrastim. Clinical trial information: 2004-001452-36, 2004-001450-84, 2004-001449-13.
An examination of crizotinib activity in human tumor primary culture micro-spheroids isolated from patients with advanced non-small cell lung cancer.
Authors: Nagourney Robert Alan , Ou Sai-Hong Ignatius , Bernard Paula J , Francisco Federico R , Evans Steven S
Published: March 2020

e13558

Background: The amino-pyridine, Crizotinib (Criz) (PF02341066, Xalkori), active against c-MET is an inhibitor of anaplastic lymphoma kinase (ALK). Identification of ALK-gene rearrangement in NSCLC led to clinical trials & FDA approval. Recognition of ROS-1 mutations as Criz targets provided additional therapy options. ALK mutations found in NSCLC also occur in lymphoma, neuro & myofibroblastic tumors but may participate in the oncogenesis of other tumors. Methods: We used ex vivo analysis of programmed cell death (EVA/PCD) (Nagourney, R. Curr Treat. Op Oncol, 2006) to examine Criz activity in human tumor 1°culture micro-spheroids, from 60 surgical specimens, with a focus on NSCLC. Using metabolic (ATP-content; mitochondrial) & morphologic (membrane integrity) endpoints, dose response curves were interpolated to LC50 values for comparison of activity by patient & tumor type. Patients were screened for ALK & ROS-1 by FISH. Results: ALK (+) tumors reveal lower LC50’s (3.4 uM) vs. ROS-1 (+) (11.5 uM), despite clinical responses in both groups. A Criz-responding patient, at 2nd biopsy for progression, reverted to Criz-resistance but developed collateral sensitivity to cytotoxics that provided durable response. Despite FISH (-), a 39 y/o nonsmoker male, revealed exquisite sensitivity to Criz by repeat EVA/PCD. At our insistence, FISH conducted at a 2nd reference lab correctly identified ALK(+) qualifying for Criz, to which he responded, now at year 2. Using low LC50 as a phenotypicmarker of Criz responsiveness, we identified activity in an extremely rare pediatric sarcoma patient. When, secondarily screened, patient found ALK (+), followed by rapid objective response to Criz. Conclusions: Primary culture analyses provide insights into Criz activity including ALK (+) vs. ROS-1(+), individual patient response profiles, and the identification of Criz candidates, who might otherwise not be screened for sensitizing mutations. By capturing human tumors in their “native” state EVA/PCD offers opportunities to study Criz for unrecognized targets and analyze novel strategies including synergy & sequence dependence, less readily examined with genomic platforms.
Preclinical toxicology studies with Y15, a novel focal adhesion kinase (FAK) inhibitor.
Authors: Cance William G. , Curtin Leslie , Buitrago Sandra , Golubovskaya Vita
Published: March 2020

e13561

Background: Focal Adhesion Kinase (FAK) is overexpressed in many types of tumors and plays a major role in survival signaling. Recently, we developed a FAK scaffolding inhibitor, Y15 that targeted the main FAK autophosphorylation site (Y397) and had high efficacy in blocking tumor growth in many xenograft mice models. We performed a small animal toxicology study of Y15 and detected the maximal tolerated dose of this drug in mice. Methods: The toxicology study was performed in CD-1 albino [Hsd:ICR(CD-1)] mice with Y15 inhibitor delivered either by intraperitoneal injection (IP) or by oral delivery. Clinical chemistry data were collected by Hemagen Analyst II automatic Chemistry analyzer. In addition, hematology, body weight, mortality, and histopathology on different organs were analyzed in all mice. Results: We delivered Y15 by IP injection at 15 (low), 30 (medium) and 45 mg/kg (high) doses daily for 5 days/week during a 28 day study, or orally by gavage at 100 and 200 mg/kg daily during a 7 day study. The maximal tolerated dose by IP during 28 day study was 30 mg/kg. The maximal tolerated dose during single oral dose administration was 200 mg/kg and 100 mg/kg during 7 day study. Y15 did not cause any mortality or statistically significant differences in the body weight at 30mg/kg IP delivery during a 28-day study and at 100 mg/kg by oral delivery during a 7 day study. There were no significant clinical chemistry changes in alkaline phosphatase, gamma glutamyltransferase, aspartate aminotransferase, alanine aminotransferase, amylase, blood urea nitrogen, glucose, phosphorus, calcium, albumin, cholesterol, creatine kinase, total bilirubin, total protein and globulin. Finally, no significant histopathological changes were observed by necropsy in different organs at 30 mg/kg by IP and at 100 mg/kg dose by oral delivery. Conclusions: The Y15 FAK autophosphorylation inhibitor is well tolerated in mice and caused no significant toxicity by IP and oral delivery, suggesting it is a promising candidate for further large animal toxicology study prior to human trials.
Pruritus in patients treated with targeted cancer therapies: Systematic review and meta-analysis.
Authors: Ensslin Courtney J. , Rosen Alyx C. , Wu Shenhong , Lacouture Mario E.
Published: March 2020

e13571

Background: Pruritus is a disabling symptom that has been anecdotally described in patients treated with targeted cancer therapies, and reports on its incidence vary. The overall risk to develop pruritus in these patients has not been systematically ascertained. We conducted a systematic review and meta-analysis of the literature to determine the incidence and risk of developing pruritus among patients treated with targeted therapies. Methods: Databases from PubMed and Web of Science from January 1998 until July 2012 and abstracts presented at the American Society of Clinical Oncology annual meetings from 2004 through 2012 were searched to identify relevant studies. The incidence and relative risk (RR) of pruritus were calculated using random-effects or fixed-effects model depending on the heterogeneity of included studies. Results: Of 5,065 studies initially identified, a total of 20,532 patients from 144 studies were included for analysis. The summary incidences of all-grade and high-grade pruritus were 17.4% (95% confidence interval (CI): 16.0%-19.0%) and 1.4% (95% CI: 1.2%-1.6%). From randomized controlled trials, patients treated with targeted therapies had a significantly increased risk of developing all-grade pruritus, with an overall RR of 2.59 (95% CI: 2.03-3.30, p<0.001); there was a significant variation among different classes of drugs (P<0.001). Conclusions: There is a significant risk of developing pruritus in cancer patients receiving targeted therapies. In order to prevent suboptimal dosing and reductions in quality of life, these patients should be counseled and treated against this disabling event.
Time-dependent estimates of recurrence and survival in colon cancer: clinical decision support system tool development for adjuvant therapy and oncological outcome assessment.
Authors: Steele Scott , Bilchik Anton , Johnson Eric , Nissan Aviram , Peoples George Earl , Eberhardt John , Kalina Philip , Petersen Benjamin , Bruecher Bjoern L. D. M. , Protic Mladjan , Avital Itzhak , Stojadinovic Alexander
Published: March 2020

e14500

Background: Unanswered questions remain regarding treatment efficacy in colon cancer (CC), especially those determining high-risk node-negative cohorts that may benefit from adjuvant therapy. We sought to evaluate the use of machine learning and classification modeling to estimate survival and recurrence in CC. Methods: We used the Department of Defense Automated Central Tumor Registry (ACTUR) to identify primary CC patients treated between January 1993 and December 2004. Cases with events or follow-up that passed quality control were stratified into one-, two-, three-, and five-year survival cohorts. ml-BBNs were trained using machine-learning algorithms and k-fold cross-validation, and receiver operating characteristic (ROC) curve analysis used for validation. Results: There were 5,301 cases stratified into cohorts. Survival cohort Areas-Under-the-Curve (AUCs) ranged from 0.85–0.90, positive-predictive-values (PPVs) for recurrence and mortality ranged from 78-84% and negative-predictive-values (NPVs) from 74-90%. Cross-validation showed that the ml-BBNs produce robust individual estimates of recurrence (p<0.001) and mortality (p<0.001) based on readily available clinical-pathological information in the context of adjuvant chemotherapy. Conclusions: Tumor registry data and machine-learned Bayesian Belief Networks produce robust classifiers. These Clinical Decision Support System tools yield clinically relevant estimates of outcomes that may assist clinicians in treatment planning.
Surgical resection of liver metastases and survival outcomes in real-life for metastatic colorectal cancer (mCRC) patients treated in front-line with cetuximab (CTX): The EREBUS cohort.
Authors: Smith Denis Michel , Sa Cunha Antonio , Rouyer Magali , Noize Pernelle , Jove Jeremy , Bignon Emmanuelle , Le Monies Alise , Mitry Emmanuel , Francois Eric , Fourrier-Réglat Annie
Published: March 2020

e14514

Background: CTX has demonstrated improved survival outcomes and resection rate in unresectable mCRC patients (pts) but little data is available from real-life use. Methods: EREBUS is a French multicentre (n=65) cohort that included pts with unresectable mCRC and wt KRAS initiating 1st-line CTX in 2009 and 2010, and followed 12 months. An expert committee validated baseline resectability and resection results. Resection rates and survival outcomes were described according to metastases site: liver-only, liver-not exclusively, other. Results: 389 pts were included: 37.8% liver-only metastases, 38.3% liver-not exclusively, 23.9% other. Baseline characteristics of the cohort: median age 64 yrs, 67.4% male, 77.9% ECOG=0-1, 55.5% primary tumor resection, 53% single metastatic site. Combined chemotherapy regimen were with irinotecan (56.6%) or oxaliplatin (38.2%). Median duration of CTX use was 4.8 months. 97 pts (24.9%, 95%CI: 20.6-29.2) had metastases resection; rates for liver-only: 36.7% (28.9-44.5), liver-not exclusively: 20.8% (14.3-27.3), other: 12.9% (6.9-21.5). Among them, 51.5% had radical resection with R0+/-radiofrequency (liver-only: 61.1% and liver-not exclusively: 32.3%), 11.3% R1+/-radiofrequency (liver-only: 16.7% and liver-not exclusively: 3.2%) and 9.4% missing metastases, 27.3%R2. 52.6% had post-operative complications (20.6% infection, 8.2% thromboembolic event, 8.2% other cardiovascular event, 3.1% death). Median PFS for the cohort was 9.7 months (8.6-10.5). In multivariate Cox analysis progression was independently associated with metastases site, liver-only: HR=1.93 (1.36-2.74), liver-not exclusively: HR=1.46 (1.06-2.02). The 1-yr OS probability was 71.3% (66.4-75.6) without any site difference. In Cox analysis, death was less likely in responders with or without metastases surgery: HR=0.26 (0.08-0.90) and HR=0.37 (0.21-0.63). Conclusions: This cohort shows that 1st-line CTX in wt KRAS mCRC allows a high proportion of metastases resection, particularly in liver-only. It also provides data on rarely studied mCRC pts (liver-not exclusively and other metastases).