Dynamic of Expression of 11 MicroRNAs in Recurrent Glioblastoma before and after Treatment with Radiotherapy and Chemotherapy

Abstract INTRODUCTION Glioblastoma is an aggressive brain tumor with a median survival of 14.6 months. We have no standard treatment for relapse and known options have limited effect. Novel treatments are necessary to improve survival and quality of life. METHODS We present our trial; phase II open label, two-armed translational study of Nivolumab and Bevacizumab for recurrent GBM, who have failed Stupp’s regimen. Patients are included in two arms depending on the possibility of salvage neurosurgical resection. Both arms receive Nivolumab and Bevacizumab administrated every second weekend, and the surgical arm also receive Nivolumab 7 days prior surgery. Forty-four patients were included by January 2021; 20 in each arm (four screen-failures). In the surgical arm, 20 fresh tumor samples as well as paired tissue from primary tumor were available. Tumor infiltrating lymphocytes (TILs) and tumor digest were produced in vitro from recurrent settings. Young TILs were expanded from fresh tumor fragments after minimal-culture, whereas rapidly expanded TILs (REP TILs) were obtained after massive expansion. By intracellular cytokine staining, we investigated the TIL reactivity after exposure to autologous tumor digest in order to evaluate whether the TILs were tumor-reactive, non-reactive or bystanders. RNA and whole exome sequencing were available before and after treatment. RESULTS Material from 19 patients was analyzed (one out of the 20 collected biopsies was limited in size, therefore no tumor digest could be produced). Four out of 19 TIL samples showed tumor reactivity after exposure to the autologous tumor digest. Tumor reactivity was ranged between 1,2 to 13,6 tox% in CD8+ TILs and between 2,8 to 10,9 tox% in CD4+ TILs. By flowcytometry we found, IgG4+ CD3+ TILS from tumor biopsies, meaning that Nivolumab were found in the brain. Currently controls are included to evaluate these results. CONCLUSIONS Updated results will be presented at SNO.

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Preliminary results using PET/MR in glioblastoma patients treated with regorafenib: an 18F-FET and DWI-ADC comparison

10.21203/rs.3.rs-296895/v1 ◽

2021 ◽

Author(s):

Alessandro Spimpolo ◽

Giuseppe Lombardi ◽

Sara Berti ◽

Cristina Campi ◽

Maria Giulia Anglani ◽

...

Keyword(s):

Early Response ◽

Recurrent Glioblastoma ◽

Response To Treatment ◽

Fet Pet ◽

Additional Information ◽

Rano Criteria ◽

Percentage Difference ◽

Before And After ◽

The Difference ◽

Cancer Network

Abstract Introduction: The use of regorafenib in recurrent glioblastoma patients has been recently approved by the Italian Medicines Agency (AIFA) and added to the National Comprehensive Cancer Network (NCCN) 2020 guidelines as a preferred regimen. Given its complex effects at the molecular level, the most appropriate imaging tools to assess early response to treatment is still a matter of debate. DWI and 18F–FET PET are promising methodologies providing additional information to the currently used RANO criteria. The aim of this study was to evaluate the variations in DWI/ADC- and 18F-FET PET-derived parameters in patients who underwent PET/MR at both baseline and soon after starting regorafenib. Method: We retrospectively selected 16 consecutive GBM patients who underwent 18F–FET PET/MR before and after two cycles of regorafenib. Patients were sorted into stable (SD) or progressive disease (PD) categories in accordance with RANO criteria. We were also able to analyze 4 SD patients who underwent a third PET/MR after another 4 cycles of regorafenib. 18F–FET uptake greater than 1.6 times the mean background activity was used to define an area to be superimposed on an ADC map at baseline and soon after treatment. A number of metrics were then derived and compared. Result: The average increases in FET and ADC pathological volumes were higher in PD than in SD patients, although in neither case did the difference reach significance. However, when the percentage difference in FET volumes was plotted against the corresponding percentage difference in ADC, a correlation was observed (R = 0.54). Patients with a twofold increase in FET after regorafenib showed a significantly higher increase in ADC pathological volume than the remaining subjects (p = 0.0023). Conclusion: In recurrent glioblastoma patients treated with regorafenib, 18F-FET and ADC metrics, being obtained from completely different measures, could serve as semi-quantitative independent biomarkers of response to treatment. These promising parameters should be tested in a larger cohort of glioblastoma patients treated with regorafenib.

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Changes of Healthy Brain Tissue after Salvage Radiotherapy of Glioblastoma Multiforme

Neuro-Oncology Advances ◽

10.1093/noajnl/vdab139 ◽

2021 ◽

Author(s):

Leandra De La Cruz ◽

Xioran Chen ◽

Ender Konugoglu ◽

I Frank Ciernik

Keyword(s):

White Matter ◽

Glioblastoma Multiforme ◽

Brain Structures ◽

Volumetric Analysis ◽

Recurrent Glioblastoma ◽

Salvage Radiotherapy ◽

Manual Segmentation ◽

Recurrent Glioblastoma Multiforme ◽

Before And After ◽

Magnetic Resonance Imaging Mri

Abstract Background Salvage radiotherapy (SRT) with photons is a valid treatment option for patients suffering from recurrent glioblastoma multiforme (GBM). However, the tolerance of healthy brain to ionizing radiation (IR) is limited. The aim of this study was to determine to what extent brain structures in the radiographically tumor-free hemisphere change after repeated radiotherapy. Methods Five of 26 patients treated with SRT for local recurrence of GBM were found to have magnetic resonance imaging (MRI) studies available for complete volumetric analysis before and after primary chemo-radiation and after SRT. Manual segmentation and joint segmentation (JS) based on a convolutional neural network were used for the segmentation of the grey matter, the white matter and the ventricles in T1 MRIs. Results Qualitative results of manual segmentation and JS were comparable. After primary chemo-radiation and SRT, the volume of the contralateral ventricles increased steadily by 1.3 to 4.75 % (SD +/- 2.8 %, R 2 = 0.82; p= <0.01) with a manual segmentation and by 1.4 to 7.4 % (SD 2.1 %, R 2 = 0.48; p= 0.025) with JS. The volume of the cortex decreased by 3.4 – 7.3 % except in one patient, the cortex volume increased by 2.5% (SD +/- 2.9%, R 2 =0.18; p = 0.19) when measured manually. When measured with JS GM decreased by 1.0 to 7.4%, in one case it increased by 3.0% (SD = 3.2%, p = 0.22, R 2= 0.18). The white matter remained stable when assessed with manual segmentation (p = 0.84, R 2 = 0.004) or JS (p = 0.44, R 2 = 0.07). Conclusion SRT of relapsed GBM leads to continuous changes of the tumor-free contralateral brain by means of manual segmentation or JS. The cortex seems more susceptible to repeated RT compared to the white matter. Larger cohort studies and complementary functional analysis are encouraged.

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Treatment of Recurrent Glioblastoma by Chronic Convection-Enhanced Delivery of Topotecan

10.1101/2021.12.04.21266935 ◽

2021 ◽

Author(s):

Eleonora F. Spinazzi ◽

Michael G. Argenziano ◽

Pavan S. Upadhyayula ◽

Matei A. Banu ◽

Justin A. Neira ◽

...

Keyword(s):

Drug Delivery ◽

Drug Distribution ◽

Response Assessment ◽

Recurrent Glioblastoma ◽

Normal Brain ◽

Convection Enhanced Delivery ◽

Before And After ◽

Treatment Response Assessment ◽

Slow Cycling ◽

Novel Drug

ABSTRACTGlioblastoma, the most common primary brain malignancy, is invariably fatal. Systemic chemotherapy is ineffective mostly because of drug delivery limitations. To overcome this, we devised an internalized pump-catheter system for direct chronic convection-enhanced delivery (CED) into peritumoral brain tissue. Topotecan (TPT) by chronic CED in 5 patients with refractory glioblastoma selectively eliminated tumor cells without toxicity to normal brain. Large, stable drug distribution volumes were non-invasively monitored with MRI of co-infused gadolinium. Analysis of multiple radiographically localized biopsies taken before and after treatment showed a decreased proliferative tumor signature resulting in a shift to a slow-cycling mesenchymal/astrocytic-like population. Tumor microenvironment analysis showed an inflammatory response and preservation of neurons. This novel drug delivery strategy and innovative clinical trial paradigm overcomes current limitations in delivery and treatment response assessment as shown here for glioblastoma and is potentially applicable for other anti-glioma agents as well as other CNS diseases.

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CTNI-20. FEASIBILITY OF SINGLE-PASS STEREOTACTIC NEEDLE BIOPSY IN RECURRENT GLIOBLASTOMA PATIENTS TO SUPPORT CLIA-CERTIFIED TUMOR PHARMACODYNAMICS FOR A PHASE 0/2 CLINICAL TRIAL

Neuro-Oncology ◽

10.1093/neuonc/noab196.245 ◽

2021 ◽

Vol 23 (Supplement_6) ◽

pp. vi63-vi63

Author(s):

Yu-Wei Chang ◽

Anita DeSantis ◽

Chelsea Pennington-Krygier ◽

Jennifer Molloy ◽

Shwetal Mehta ◽

...

Keyword(s):

Clinical Trial ◽

Needle Biopsy ◽

Stereotactic Biopsy ◽

Recurrent Glioblastoma ◽

Experimental Therapy ◽

Time Interval ◽

Single Pass ◽

Phase 0 ◽

Before And After ◽

Pre Treatment

Abstract BACKGROUND In neuro-oncology clinical trials, pharmacodynamic (PD) analysis of tumor tissue before and after experimental therapy is challenging. Today, Phase 0 studies in brain tumor patients typically employ archival tissue from a prior resection as the baseline PD comparator. However, the time-interval between tissue acquisitions can be months to years and often includes confounding exposure to other therapies. Here, we demonstrate the feasibility of a pre-treatment, single-pass stereotactic needle biopsy to support CLIA-certified genetic screening and tumor PD analysis in a Phase 0/2 clinical trial. METHODS In support of an ongoing Phase 0/2 ‘trigger’ trial testing CDK4/6 plus ERK1/2 inhibition in recurrent glioblastoma (GBM), a single-pass stereotactic biopsy was completed in select patients prior to enrollment. Each biopsy yields six tissue cores for immunohistochemistry (IHC) and genetic characterization. Using CLIA-certified protocols, genomic DNA (gDNA) was extracted from two cores, followed by library preparation and next-generation sequencing of a customized panel of 37 genes (IvySeq). In the four remaining cores, IHC staining was performed for RB, pRB, pERK, pS6, CDKN2A, MIB-1, ClCas-3 and pH2AX. RESULTS Single-pass stereotactic needle biopsies were collected from five recurrent GBM patients. No intraoperative, perioperative, or radiographic evidence of morbidity was observed. 800-2400ng of gDNA was isolated from each stereotactic biopsy. IvySeq analysis detected CDKN2A deletion, ATRX loss, and mutations in IDH, PTEN and TP53 genes. Based on IHC and genetic analyses of the biopsied samples, three patients were enrolled into the Phase 0/2 clinical trial and baseline value were used for PD analysis. CONCLUSION Single-pass stereotactic needle biopsy is a feasible and safe strategy to collect pre-treatment tissue in support of a Phase 0 clinical trial, enabling CLIA-certified genetic analysis for trial screening and tumor PD analysis.

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Eosinophil and lymphocyte counts predict bevacizumab response and survival in recurrent glioblastoma

Neuro-Oncology Advances ◽

10.1093/noajnl/vdaa031 ◽

2020 ◽

Vol 2 (1) ◽

Author(s):

Eugene J Vaios ◽

Sebastian F Winter ◽

Alona Muzikansky ◽

Brian V Nahed ◽

Jorg Dietrich

Keyword(s):

Multivariate Analysis ◽

Treatment Response ◽

Clinical Outcomes ◽

Survival Probability ◽

Progression Free Survival ◽

Recurrent Glioblastoma ◽

Platelet Counts ◽

Before And After ◽

Complete Blood Counts ◽

Recurrent Gbm

Abstract Background There is a lack of biomarkers to identify glioblastoma (GBM) patients who may benefit from specific salvage therapies, such as the anti-angiogenic agent bevacizumab. We hypothesized that circulating blood counts may serve as biomarkers for treatment response and clinical outcomes. Methods Complete blood counts, clinical data, and radiographic information were collected retrospectively from 84 recurrent GBM patients receiving bevacizumab (10 mg/kg every 2 weeks). Significant biomarkers were categorized into quartiles and the association with clinical outcomes was assessed using the Kaplan–Meier method. Results The median treatment duration and survival on bevacizumab (OS-A) was 88 and 192 days, respectively. On multivariate analysis, MGMT promoter methylation (hazard ratio [HR] 0.504, P = .031), increases in red blood cells (HR 0.496, P = .035), and increases in eosinophils (HR 0.048, P = .054) during treatment predicted improved OS-A. Patients in the first and fourth quartiles of eosinophil changes had a 12-month survival probability of 5.6% and 41.2% (P < .0001), respectively. Treatment response was associated with increases in eosinophil counts (P = .009) and improved progression-free survival (P = .013). On multivariate analysis, increases in lymphocyte counts among responders predicted improved OS-A (HR 0.389, P = .044). Responders in the first and fourth quartiles of lymphocyte changes had a 12-month survival probability of 0% and 44.4% (P = .019), respectively. Changes in platelet counts differed before and after radiographic response (P = .014). Conclusions Changes in circulating eosinophil, lymphocyte, and platelet counts may predict treatment response and clinical outcomes in patients with recurrent GBM receiving bevacizumab.

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Recurrent glioblastoma: Stratification of patient survival using tumor volume before and after antiangiogenic treatment.

Journal of Clinical Oncology ◽

10.1200/jco.2013.31.15_suppl.2075 ◽

2013 ◽

Vol 31 (15_suppl) ◽

pp. 2075-2075 ◽

Cited By ~ 2

Author(s):

Raymond Huang ◽

Rifaquat Rahman ◽

Alhafidz Hamdan ◽

Caroline Kane ◽

Christina Chen ◽

...

Keyword(s):

Overall Survival ◽

Tumor Volume ◽

Proportional Hazards ◽

Proportional Hazards Model ◽

Progression Free Survival ◽

Cox Proportional Hazards ◽

Recurrent Glioblastoma ◽

Cox Proportional Hazards Model ◽

Percentage Change ◽

Before And After

2075 Background: Although antiangiogenic therapies can induce a dramatic clinical and imaging response in patients with recurrent glioblastoma, their benefit on overall survival is less pronounced. We assessed whether tumor volume based on magnetic resonance imaging (MRI) before and after treatment with bevacizumab can stratify patients in terms of progression-free survival (PFS) and overall survival (OS). Methods: Baseline and post-treatment MRI scans of 93 patients with recurrent glioblastoma treated with bevacizumab were evaluated for volume of the enhancing tumor, as well as volume of the T2/FLAIR hyperintensity. The Cox proportional hazards model was used in univariate and multivariate settings to identify volume parameters prognostic and predictive for PFS and OS. Results: Our results show that baseline enhancing tumor volume were predictive of PFS (p = 0.047) and OS (p = 0.011). Specifically, patients with pretreatment enhancing tumor volume less than 18 cm3 have a median OS of 45 weeks, while patients with tumor volume greater than 18 cm3 have a median OS of 26 weeks. In addition, enhancing tumor volume 3 to 6 weeks after treatment, as well as percentage change in volume, are independent predictors of PFS and OS. Neither T2/FLAIR volume nor percentage change in T2/FLAIR volume was associated with survival. Conclusions: These results indicate that measurement of enhancing tumor volume can help identify patients more likely to benefit from bevacizumab therapy.

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Deformation Replication

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100068394 ◽

1970 ◽

Vol 28 ◽

pp. 280-281

Author(s):

J. Temple Black

Keyword(s):

Cutting Speed ◽

Machining Process ◽

Depth Of Cut ◽

Rake Face ◽

Orthogonal Machining ◽

Aluminum Chips ◽

Before And After ◽

Materials Used ◽

Glass Knife ◽

Very High

Tool materials used in ultramicrotomy are glass, developed by Latta and Hartmann (1) and diamond, introduced by Fernandez-Moran (2). While diamonds produce more good sections per knife edge than glass, they are expensive; require careful mounting and handling; and are time consuming to clean before and after usage, purchase from vendors (3-6 months waiting time), and regrind. Glass offers an easily accessible, inexpensive material ($0.04 per knife) with very high compressive strength (3) that can be employed in microtomy of metals (4) as well as biological materials. When the orthogonal machining process is being studied, glass offers additional advantages. Sections of metal or plastic can be dried down on the rake face, coated with Au-Pd, and examined directly in the SEM with no additional handling (5). Figure 1 shows aluminum chips microtomed with a 75° glass knife at a cutting speed of 1 mm/sec with a depth of cut of 1000 Å lying on the rake face of the knife.

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The Clinical, Roentgenological and Morphological Effects of an Acute Exposure of Phosgene on the Lungs of Dogs

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100065237 ◽

1971 ◽

Vol 29 ◽

pp. 236-237

Author(s):

R. F. Bils ◽

W. F. Diller ◽

F. Huth

Keyword(s):

Latent Period ◽

Chemical Industry ◽

Toxic Substance ◽

Lung Edema ◽

X Rays ◽

Beagle Dogs ◽

Male And Female ◽

Morphological Alterations ◽

Before And After ◽

Electron Microscopes

Phosgene still plays an important role as a toxic substance in the chemical industry. Thiess (1968) recently reported observations on numerous cases of phosgene poisoning. A serious difficulty in the clinical handling of phosgene poisoning cases is a relatively long latent period, up to 12 hours, with no obvious signs of severity. At about 12 hours heavy lung edema appears suddenly, however changes can be seen in routine X-rays taken after only a few hours' exposure (Diller et al., 1969). This study was undertaken to correlate these early changes seen by the roengenologist with morphological alterations in the lungs seen in the'light and electron microscopes.Forty-two adult male and female Beagle dogs were selected for these exposure experiments. Treated animals were exposed to 94.5-107-5 ppm phosgene for 10 min. in a 15 m3 chamber. Roentgenograms were made of the thorax of each animal before and after exposure, up to 24 hrs.

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Ultrastructure of Melanin Formation in Curvularia sp., Alternaria sp., and Drechslera Sorokiniana

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100079449 ◽

1977 ◽

Vol 35 ◽

pp. 398-399

Author(s):

M. H. Wheeler ◽

W. J. Tolmsoff ◽

A. A. Bell

Keyword(s):

Potassium Phosphate ◽

Thielaviopsis Basicola ◽

Wild Type ◽

Potassium Phosphate Buffer ◽

Transmission Microscopy ◽

Electron Transmission ◽

Melanin Formation ◽

Before And After ◽

Alternaria Sp ◽

Electron Transmission Microscopy

(+)-Scytalone [3,4-dihydro-3,6,8-trihydroxy-l-(2Hj-naphthalenone] and 1,8-di- hydroxynaphthalene (DHN) have been proposed as intermediates of melanin synthesis in the fungi Verticillium dahliae (1, 2, 3, 4) and Thielaviopsis basicola (4, 5). Scytalone is enzymatically dehydrated by V. dahliae to 1,3,8-trihydroxynaphthalene which is then reduced to (-)-vermelone [(-)-3,4- dihydro-3,8-dihydroxy-1(2H)-naphthalenone]. Vermelone is subsequently dehydrated to DHN which is enzymatically polymerized to melanin.Melanin formation in Curvularia sp., Alternaria sp., and Drechslera soro- kiniana was examined by light and electron-transmission microscopy. Wild-type isolates of each fungus were compared with albino mutants before and after treatment with 1 mM scytalone or 0.1 mM DHN in 50 mM potassium phosphate buffer, pH 7.0. Both chemicals were converted to dark pigments in the walls of hyphae and conidia of the albino mutants. The darkened cells were similar in appearance to corresponding cells of the wild types under the light microscope.

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