Resonancia intraoperatoria de alto campo: cómo optimizar su uso en nuestro modelo sanitario / Intraoperative high-field resonance: how to optimize its use in our healthcare system

Antecedentes y objetivo: La RM intraoperatoria (RMio) consiste en la realización de una resonancia durante la cirugía de una lesión cerebral o espinal. Es una técnica segura y útil, aunque está disponible en pocos hospitales y algunos aspectos no están perfectamente definidos ni estandarizados, por lo que cada centro elabora sus propias soluciones. Nuestro objetivo es describir la técnica utilizada para la realización de RMio, evaluar los cambios que se han realizado para optimizar su uso desde el comienzo y facilitar la puesta en marcha de una resonancia intraoperatoria en otros departamentos de neurocirugía. Material y métodos: Estudio prospectivo de pacientes intervenidos consecutivamente con RMio, describiendo el tipo de tumor, datos clínicos, tiempo y secuencias de RMio, empleo de neurofisiología intraoperatoria, volumen tumoral preoperatorio, tras la RMio, y postoperatorio, y complicaciones observadas. Resultados: Se realizó RMio en 38 pacientes seleccionados de los 425 tumores cerebrales (9%) operados en este intervalo. Los tipos tumorales fueron: 11 glioblastomas, 8 astrocitomas anaplásicos, 5 astrocitomas difusos, 4 meningiomas, 3 oligodendrogliomas, 2 metástasis, 2 quistes epidermoides, 1 astroblastoma, 1 quiste aracnoideo y 1 adenoma hipofisario.La edad media fue de 45 años. El volumen tumoral preoperatorio medio fue 45,22 cc, tras la RMio de 5,08 cc y el postoperatorio de 1,28 cc.En el 76% se amplió la resección tras la RMio. En 15 pacientes se consiguió una resección completa y en 8 se objetivó un resto menor de 1cc. En 13 pacientes se dejó un resto intencional en área elocuente o regiones de base de cráneo (volumen medio 7cc).En un 5% se detectaron complicaciones de sangrado e isquemia de forma precoz en la RMio.La realización de la RMio requirió una media de 47 minutos...(AU)

Background and aims: Intraoperative MRI (ioMRI) consists of performing a MRI during brain or spinal surgery. Although it is a safe and useful technique, it is available in a few hospitals. This means some aspects are not perfectly defined or standardized, forcing each center to develop its own solutions. Our goal is to describe the technique, evaluate the changes made to optimize its use and thus be able to facilitate the intraoperative resonance implementation in other neurosurgery departments. Methods: A prospective analysis of patients consecutively operated using high-field ioMRI guidance was carried out, describing the type of tumor, clinical data, time and sequences of ioMR, use of intraoperative neurophysiology, preoperative tumor volume, after ioMR, and postoperative, as well as complications. Resultsio: MR was performed in 38 patients selected from among 425 brain tumors (9%) operated on in this interval. The tumor types were: 11 glioblastomas, 8 anaplastic astrocytomas, 5 diffuse astrocytomas, 4 meningiomas, 3 oligodendrogliomas, 2 metastases, 2 epidermoid cysts, 1 astroblastoma, 1 arachnoid cyst and 1 pituitary adenoma.The mean age was 45 years. The mean preoperative tumor volume was 45.22cc, after the ioMR 5.08cc and postoperative 1.28cc. Resection was extended after ioMR in 76%. Gross total resection was achieved in 15 patients and residual tumor of less than 1cc was observed in 8. An intentional tumor tissue was left in an eloquent brain region (mean volume 7cc) in 13 patients.Bleeding and ischemia complications were detected early on ioMR in 5%.MRI length was 47minutes on average. Conclusions: Intraoperative MRI was a useful and safe technique, and no associated complications were registered. (AU)

Oncolytic DNX-2401 Virus for Pediatric Diffuse Intrinsic Pontine Glioma.

BACKGROUND: Pediatric patients with diffuse intrinsic pontine glioma (DIPG) have a poor prognosis, with a median survival of less than 1 year. Oncolytic viral therapy has been evaluated in patients with pediatric gliomas elsewhere in the brain, but data regarding oncolytic viral therapy in patients with DIPG are lacking. METHODS: We conducted a single-center, dose-escalation study of DNX-2401, an oncolytic adenovirus that selectively replicates in tumor cells, in patients with newly diagnosed DIPG. The patients received a single virus infusion through a catheter placed in the cerebellar peduncle, followed by radiotherapy. The primary objective was to assess the safety and adverse-event profile of DNX-2401. The secondary objectives were to evaluate the effect of DNX-2401 on overall survival and quality of life, to determine the percentage of patients who have an objective response, and to collect tumor-biopsy and peripheral-blood samples for correlative studies of the molecular features of DIPG and antitumor immune responses. RESULTS: A total of 12 patients, 3 to 18 years of age, with newly diagnosed DIPG received 1×1010 (the first 4 patients) or 5×1010 (the subsequent 8 patients) viral particles of DNX-2401, and 11 received subsequent radiotherapy. Adverse events among the patients included headache, nausea, vomiting, and fatigue. Hemiparesis and tetraparesis developed in 1 patient each. Over a median follow-up of 17.8 months (range, 5.9 to 33.5), a reduction in tumor size, as assessed on magnetic resonance imaging, was reported in 9 patients, a partial response in 3 patients, and stable disease in 8 patients. The median survival was 17.8 months. Two patients were alive at the time of preparation of the current report, 1 of whom was free of tumor progression at 38 months. Examination of a tumor sample obtained during autopsy from 1 patient and peripheral-blood studies revealed alteration of the tumor microenvironment and T-cell repertoire. CONCLUSIONS: Intratumoral infusion of oncolytic virus DNX-2401 followed by radiotherapy in pediatric patients with DIPG resulted in changes in T-cell activity and a reduction in or stabilization of tumor size in some patients but was associated with adverse events. (Funded by the European Research Council under the European Union's Horizon 2020 Research and Innovation Program and others; EudraCT number, 2016-001577-33; ClinicalTrials.gov number, NCT03178032.).

Intraoperative high-field resonance: How to optimize its use in our healthcare system.

BACKGROUND AND AIMS: Intraoperative MRI (ioMRI) consists of performing a MRI during brain or spinal surgery. Although it is a safe and useful technique, it is available in a few hospitals. This means some aspects are not perfectly defined or standardized, forcing each center to develop its own solutions. Our goal is to describe the technique, evaluate the changes made to optimize its use and thus be able to facilitate the intraoperative resonance implementation in other neurosurgery departments. METHODS: A prospective analysis of patients consecutively operated using high-field ioMRI guidance was carried out, describing the type of tumor, clinical data, time and sequences of ioMR, use of intraoperative neurophysiology, preoperative tumor volume, after ioMR, and postoperative, as well as complications. RESULTS: ioMR was performed in 38 patients selected from among 425 brain tumors (9%) operated on in this interval. The tumor types were: 11 glioblastomas, 8 anaplastic astrocytomas, 5 diffuse astrocytomas, 4 meningiomas, 3 oligodendrogliomas, 2 metastases, 2 epidermoid cysts, 1 astroblastoma, 1 arachnoid cyst and 1 pituitary adenoma. The mean age was 45 years. The mean preoperative tumor volume was 45.22cc, after the ioMR 5.08cc and postoperative 1.28cc. Resection was extended after ioMR in 76%. Gross total resection was achieved in 15 patients and residual tumor of less than 1cc was observed in 8. An intentional tumor tissue was left in an eloquent brain region (mean volume 7cc) in 13 patients. Bleeding and ischemia complications were detected early on ioMR in 5%. MRI length was 47 min on average. CONCLUSIONS: Intraoperative MRI was a useful and safe technique, and no associated complications were registered.

Neurosurgical emergency management during the lockdown period in health care regions in Spain with different COVID-19 impact: lessons learned to improve outcomes on the future waves.

BACKGROUND: COVID-19 has overloaded health care systems, testing the capacity and response in every European region. Concerns were raised regarding the impact of resources' reorganization on certain emergency pathology management. The aim of the present study was to assess the impact of the outbreak (in terms of reduction of neurosurgical emergencies) during lockdown in different regions of Spain. METHODS: We analyzed the impact of the outbreak in four different affected regions by descriptive statistics and univariate comparison with same period of two previous years. These regions differed in their incidence level (high/low) and in the time of excess mortality with respect to lockdown declaration. That allowed us to analyze their influence on the characteristics of neurosurgical emergencies registered for every region. RESULTS: 1185 patients from 18 neurosurgical centers were included. Neurosurgical emergencies that underwent surgery dropped 24.41% and 28.15% in 2020 when compared with 2019 and 2018, respectively. A higher reduction was reported for the most affected regions by COVID-19. Non-traumatic spine experienced the most significant decrease in number of cases. Life-threatening conditions did not suffer a reduction in any health care region. CONCLUSIONS: COVID-19 affected dramatically the neurosurgical emergency management. The most significant reduction in neurosurgical emergencies occurred on those regions that were hit unexpectedly by the pandemic, as resources were focused on fighting the virus. As a consequence, life-threating and non-life-threatening conditions' mortality raised. Results in regions who had time to prepare for the hit were congruent with an organized and sensible neurosurgical decision-making.

Ki-67 Proliferative Activity in the Tumor Margins as a Robust Prognosis Factor in Glioblastoma Patients.

INTRODUCTION: The infiltrative margin of glioblastomas (GBM) contains proliferative tumor cells difficult to estimate radiologically as they are included in the hyperintense signal of T2 sequences and they remain in the cavity margin after tumor resection. The amount of these cells could determine overall survival (OS) of these patients. MATERIAL AND METHODS: From October 2007 to January 2010, patients whose MRI were suggestive of newly diagnosed, resectable high-grade glioma were operated using fluorescence-guided surgery (FGS). Separate samples were selectively taken from nonfluorescent white matter areas just adjacent to the border of the pale fluorescence and staining was made for Ki-67. OS was analyzed with Kaplan-Meier and Cox regression. Multivariate analysis included the following prognosis variables: age, extent of resection (EOR), O-6-methylguanine-DNA methyltransferase (MGMT) promoter methylation, and performance status index. RESULTS: Sample included 65 patients, comprising 37 men and 28 women, with a median Karnofsky Performance Score (KPS) of 80 (40-100) and mean age of 60 (34-78) years. Mean preoperative tumor volume was 35.8 mL. EOR was 100% in 52 patients (80%), with the lower EOR being 88%. For Ki-67, 39 patients had <5% and 26 had ≥5%. OS was 26.8 months (95% confidence interval [CI]: 18.9-28.2) for the Ki-67 low group versus 15.8 months (95% CI: 7.7-18.2) for the Ki-67 high group (p = 0.002). CONCLUSION: Proliferative activity in the normal-looking brain around the resection cavity measured with Ki-67 immunostaining is an important independent prognostic factor for GBM cases with complete resection of enhancing tumor. When complete resection is not reached, this factor is not relevant for prognosis.

Biopsy in diffuse pontine gliomas: expert neurosurgeon opinion-a survey from the SIOPE brain tumor group.

INTRODUCTION: The prognosis of diffuse intrinsic pontine glioma (DIPG) is poor. The role of biopsy in DIPG remains controversial since the diagnosis may be established with imaging alone. Recent advances in understanding molecular biology and targeting of brain tumors have created a renewed interest in biopsy for DIPG. The Neurosurgery Working Group (NWG) of the SIOP-Europe Brain Tumor Group (BTG) undertook a survey among international pediatric neurosurgeons to define their current perceptions and practice regarding DIPG biopsy. METHODS: The NWG developed a 20-question survey which was emailed to neurosurgeons in the International Society for Pediatric Neurosurgery (ISPN). The questionnaire included questions on diagnosis, indications, and techniques for biopsy, clinical trials, and healthcare infrastructure. RESULTS: The survey was sent to 202 neurosurgeons and 73 (36%) responded. Consensus of > 75% agreement was reached for 12/20 questions, which included (1) radiological diagnosis of DIPG is sufficient outside a trial, (2) clinical trial-based DIPG biopsy is justified if molecular targets are investigated and may be used for treatment, and (3) morbidity/mortality data must be collected to define the risk:benefit ratio. The remaining 8/20 questions proved controversial and failed to reach consensus. CONCLUSIONS: Routine DIPG biopsy continues to be debated. Most neurosurgeons agreed that DIPG biopsy within a clinical trial should be supported, with the aims of defining the procedure risks, improving understanding of tumor biology, and evaluating new treatment targets. Careful family counseling and consent remain important.

Correction: Hypofractionated radiation therapy and temozolomide in patients with glioblastoma and poor prognostic factors. A prospective, single-institution experience.

[This corrects the article DOI: 10.1371/journal.pone.0217881.].

Hypofractionated radiation therapy and temozolomide in patients with glioblastoma and poor prognostic factors. A prospective, single-institution experience.

BACKGROUND: Hypofractionated radiation therapy is a feasible and safe treatment option in elderly and frail patients with glioblastoma. The aim of this study was to evaluate the effectiveness of hypofractionated radiation therapy with concurrent temozolomide in terms of feasibility and disease control in primary glioblastoma patients with poor prognostic factors other than advanced age, such as post-surgical neurological complications, high tumor burden, unresectable or multifocal lesions, and potential low treatment compliance due to social factors or rapidly progressive disease. MATERIAL AND METHODS: GTV included the surgical cavity plus disease visible in T1WI-MRI, FLAIR-MRI and in the MET-uptake. The CTV was defined as the GTV plus 1.5-2 cm margin; the PTV was the CTV+0.3 cm margin. Forty, fourty-five, and fifty grays in 15 fractions were prescribed to 95% of PTV, CTV, and GTV, respectively. Treatment was delivered using IMRT or the VMAT technique. Simultaneously, 75 mg/m2/day of temozolomide were administered. RESULTS: Between January 2010 and November 2017, we treated a total of 17 patients. The median age at diagnosis was 68-years; median KPS was 50-70%. MGMT-methylation status was negative in 5 patients, and 8 patients were IDH-wildtype. Eight of 18 patients were younger than 65-years. Median tumor volume was 26.95cc; median PTV volume was 322cc. Four lesions were unresectable; 6 patients underwent complete surgical resection. Median residual volume was 1.14cc. Progression-free survival was 60% at 6 months, 33% at 1-year and 13% at 2-years (median OS = 7 months). No acute grade 3-5 toxicities were documented. Symptomatic grade 3 radiation necrosis was observed in one patient. CONCLUSIONS: Patients with poor clinical factors other than advanced age can be selected for hypofractionated radiotherapy. The OS and PFS rates obtained in our series are similar to those in patients treated with standard fractionation, assuring good treatment adherence, low rates of toxicity and probable improved cost-effectiveness.

Delta-24-RGD combined with radiotherapy exerts a potent antitumor effect in diffuse intrinsic pontine glioma and pediatric high grade glioma models.

Pediatric high grade gliomas (pHGG), including diffuse intrinsic pontine gliomas (DIPGs), are aggressive tumors with a dismal outcome. Radiotherapy (RT) is part of the standard of care of these tumors; however, radiotherapy only leads to a transient clinical improvement. Delta-24-RGD is a genetically engineered tumor-selective adenovirus that has shown safety and clinical efficacy in adults with recurrent gliomas. In this work, we evaluated the feasibility, safety and therapeutic efficacy of Delta-24-RGD in combination with radiotherapy in pHGGs and DIPGs models. Our results showed that the combination of Delta-24-RGD with radiotherapy was feasible and resulted in a synergistic anti-glioma effect in vitro and in vivo in pHGG and DIPG models. Interestingly, Delta-24-RGD treatment led to the downregulation of relevant DNA damage repair proteins, further sensitizing tumors cells to the effect of radiotherapy. Additionally, Delta-24-RGD/radiotherapy treatment significantly increased the trafficking of immune cells (CD3, CD4+ and CD8+) to the tumor niche compared with single treatments. In summary, administration of the Delta-24-RGD/radiotherapy combination to pHGG and DIPG models is safe and significantly increases the overall survival of mice bearing these tumors. Our data offer a rationale for the combination Delta-24-RGD/radiotherapy as a therapeutic option for children with these tumors. SIGNIFICANCE: Delta-24-RGD/radiotherapy administration is safe and significantly increases the survival of treated mice. These positive data underscore the urge to translate this approach to the clinical treatment of children with pHGG and DIPGs.

Neoadjuvant nivolumab modifies the tumor immune microenvironment in resectable glioblastoma.

Glioblastoma is the most common primary central nervous system malignancy and has a poor prognosis. Standard first-line treatment, which includes surgery followed by adjuvant radio-chemotherapy, produces only modest benefits to survival1,2. Here, to explore the feasibility, safety and immunobiological effects of PD-1 blockade in patients undergoing surgery for glioblastoma, we conducted a single-arm phase II clinical trial (NCT02550249) in which we tested a presurgical dose of nivolumab followed by postsurgical nivolumab until disease progression or unacceptable toxicity in 30 patients (27 salvage surgeries for recurrent cases and 3 cases of primary surgery for newly diagnosed patients). Availability of tumor tissue pre- and post-nivolumab dosing and from additional patients who did not receive nivolumab allowed the evaluation of changes in the tumor immune microenvironment using multiple molecular and cellular analyses. Neoadjuvant nivolumab resulted in enhanced expression of chemokine transcripts, higher immune cell infiltration and augmented TCR clonal diversity among tumor-infiltrating T lymphocytes, supporting a local immunomodulatory effect of treatment. Although no obvious clinical benefit was substantiated following salvage surgery, two of the three patients treated with nivolumab before and after primary surgery remain alive 33 and 28 months later.

Management of diffuse glioma.

Diffuse gliomas constitute a diverse group of malignant brain tumors with varying aggressive course and heterogeneous survival. Although the mainstay of treatment of these distinct tumors is still based on the combination of surgery and classical therapeutic weapons such as radiotherapy and chemotherapy, important advances have been achieved over the past decades leading to meaningful improvements in survival times. In addition, recent progress in molecular profiling has allowed the identification of patients with better prognosis and more likely to respond to specific antitumor treatment. This is particularly true for grade II and III 1p/19q-codeleted gliomas, a subset of tumors in which data maturation after long-term follow-up have proved extremely important for accurate assessment of efficacy. This article aims at providing a review of the current specific antitumor treatment of diffuse glioma, particularly grade II and III glioma and glioblastoma, with special emphasis on the most relevant clinical trials conducted in these populations of patients.

Rizotomía dorsal selectiva: revisión bibliográfica de esta técnica para el tratamiento de la espasticidad en la parálisis cerebral infantil / Selective dorsal rhizotomy: a review of the literature on this technique for the treatment of spasticity in infantile cerebral palsy

Introducción. La parálisis cerebral infantil conforma una entidad bien conocida cuya prevalencia ha variado poco a lo largo de los años. El subtipo más común es la diplejía espástica, y la espasticidad es el síntoma más incapacitante. Para su tratamiento es preciso una intervención multidisciplinar que aúna rehabilitación, uso de fármacos y cirugía ortopédica y del sistema nervioso, donde destaca la rizotomía dorsal selectiva. Objetivo. Mostrar una amplia revisión del uso, indicaciones y consecuencias a largo plazo de la rizotomía dorsal selectiva. Desarrollo. Se trata de una intervención mínimamente invasiva dirigida a disminuir la espasticidad en las extremidades inferiores con el fin de mejorar la deambulación, aminorar el dolor, facilitar los cuidados de la vida diaria y reducir la necesidad de cirugías ortopédicas. En la bibliografía hay una amplia variabilidad de criterios para su uso, y la principal indicación es la diplejía espástica con ausencia de distonía. Diversos países lo practican de forma rutinaria, mientras que en el nuestro no tenemos aún constancia de ello. Conclusiones. Tras la revisión de la bibliografía consideramos que existe suficiente experiencia como para afirmar que la rizotomía dorsal selectiva es una técnica segura y sencilla de la que muchos pacientes con espasticidad de las extremidades inferiores secundaria a parálisis cerebral infantil se pueden beneficiar a corto y largo plazo

Introduction. Infantile cerebral palsy is a well-known condition, the prevalence of which has varied only slightly over the years. The most common subtype is spastic diplegia, and spasticity is the most disabling symptom. Its treatment involves a multidisciplinary intervention that includes rehabilitation, the use of drugs, and orthopaedic and nervous system surgery, where selective dorsal rhizotomy is a prominent procedure. Aim. To present a thorough review of the use, indication and long-term consequences of selective dorsal rhizotomy. Development. It is a minimally invasive procedure aimed at reducing spasticity in the lower extremities in order to improve the ability to walk, lessen pain, facilitate care in everyday life and diminish the need for orthopaedic surgery. The literature contains a wide range of criteria for its use, and the main indication is spastic diplegia with the absence of dystonia. It is routinely performed in several countries, while we have no evidence of its application in ours. Conclusions. Following the literature review, we believe there is enough experience to state that selective dorsal rhizotomy is a safe and simple technique from which many patients with spasticity of the lower limbs secondary to infantile cerebral palsy can benefit in both the short and the long term

DNX-2401, an Oncolytic Virus, for the Treatment of Newly Diagnosed Diffuse Intrinsic Pontine Gliomas: A Case Report.

Diffuse intrinsic pontine gliomas (DIPGs) are aggressive glial brain tumors that primarily affect children, for which there is no curative treatment. Median overall survival is only one year. Currently, the scientific focus is on expanding the knowledge base of the molecular biology of DIPG, and identifying effective therapies. Oncolytic adenovirus DNX-2401 is a replication-competent, genetically modified virus capable of infecting and killing glioma cells, and stimulating an anti-tumor immune response. Clinical trials evaluating intratumoral DNX-2401 in adults with recurrent glioblastoma have demonstrated that the virus has a favorable safety profile and can prolong survival. Subsequently, these results have encouraged the transition of this biologically active therapy from adults into the pediatric population. To this aim, we have designed a clinical Phase I trial for newly diagnosed pediatric DIPG to investigate the feasibility, safety, and preliminary efficacy of delivering DNX-2401 into tumors within the pons following biopsy. This case report presents a pediatric patient enrolled in this ongoing Phase I trial for children and adolescents with newly diagnosed DIPG. The case involves an 8-year-old female patient with radiologically diagnosed DIPG who underwent stereotactic tumor biopsy immediately followed by intratumoral DNX-2401 in the same biopsy track. Because there were no safety concerns or new neurological deficits, the patient was discharged 3 days after the procedures. To our knowledge, this is the first report of intratumoral DNX-2401 for a patient with DIPG in a clinical trial. We plan to demonstrate that intratumoral delivery of an oncolytic virus following tumor biopsy for pediatric patients with DIPG is a novel and feasible approach and that DNX-2401 represents an innovative treatment for the disease.

The impact of intraoperative magnetic resonance in routine pediatric neurosurgical practice-a 6-year appraisal.

BACKGROUND: The intraoperative magnetic resonance scanner (ioMR) was introduced in our unit in 2009, and has been used routinely since then. OBJECTIVE: This study aims to describe indications, radiological features, and clinical outcomes of the patients operated on with ioMRI and analyze our experience. METHODS: A retrospective analysis of a prospective surgical database has been performed, including surgical procedure, intent, radiological reports, need for second-look surgery, and complications, supplemented by further review of the clinical notes and the scans. RESULTS: From 2009 to 2015, 255 surgical procedures with ioMR were performed: 175 were craniotomies for tumor excision, 65 were epilepsy related, and 15 were biopsies or cyst drainages. The mean age was 9.4 years. One ioMR was performed in 79.5% patients; the mean duration of the MR was 41 min. In 172 cases (67.4%), no actions followed the ioMR. When the aim of the surgery was debulking of the tumor, the percentage of patients in which the ioMR was followed by resection was higher than when complete resection was the aim (56 vs 27.5%). The complication rate was not increased when compared with our previous results (infection 1%, neurological deficits 12%). CONCLUSION: This is the largest published series of ioMRI-aided pediatric neurosurgery to date. We have demonstrated that it can be used safely and routinely in pediatric neurosurgical procedures at any age, assisting the surgeon in achieving the best extent of resection and aiding in intra-operative decision-making for tumor- and non-tumor-related intracranial pathology.

Phase I Study of DNX-2401 (Delta-24-RGD) Oncolytic Adenovirus: Replication and Immunotherapeutic Effects in Recurrent Malignant Glioma.

Purpose DNX-2401 (Delta-24-RGD; tasadenoturev) is a tumor-selective, replication-competent oncolytic adenovirus. Preclinical studies demonstrated antiglioma efficacy, but the effects and mechanisms of action have not been evaluated in patients. Methods A phase I, dose-escalation, biologic-end-point clinical trial of DNX-2401 was conducted in 37 patients with recurrent malignant glioma. Patients received a single intratumoral injection of DNX-2401 into biopsy-confirmed recurrent tumor to evaluate safety and response across eight dose levels (group A). To investigate the mechanism of action, a second group of patients (group B) underwent intratumoral injection through a permanently implanted catheter, followed 14 days later by en bloc resection to acquire post-treatment specimens. Results In group A (n = 25), 20% of patients survived > 3 years from treatment, and three patients had a ≥ 95% reduction in the enhancing tumor (12%), with all three of these dramatic responses resulting in > 3 years of progression-free survival from the time of treatment. Analyses of post-treatment surgical specimens (group B, n = 12) showed that DNX-2401 replicates and spreads within the tumor, documenting direct virus-induced oncolysis in patients. In addition to radiographic signs of inflammation, histopathologic examination of immune markers in post-treatment specimens showed tumor infiltration by CD8+ and T-bet+ cells, and transmembrane immunoglobulin mucin-3 downregulation after treatment. Analyses of patient-derived cell lines for damage-associated molecular patterns revealed induction of immunogenic cell death in tumor cells after DNX-2401 administration. Conclusion Treatment with DNX-2401 resulted in dramatic responses with long-term survival in recurrent high-grade gliomas that are probably due to direct oncolytic effects of the virus followed by elicitation of an immune-mediated antiglioma response.

The aberrant splicing of BAF45d links splicing regulation and transcription in glioblastoma.

Background: Glioblastoma, the most aggressive primary brain tumor, is genetically heterogeneous. Alternative splicing (AS) plays a key role in numerous pathologies, including cancer. The objectives of our study were to determine whether aberrant AS could play a role in the malignant phenotype of glioma and to understand the mechanism underlying its aberrant regulation. Methods: We obtained surgical samples from patients with glioblastoma who underwent 5-aminolevulinic fluorescence-guided surgery. Biopsies were taken from the tumor center as well as from adjacent normal-appearing tissue. We used a global splicing array to identify candidate genes aberrantly spliced in these glioblastoma samples. Mechanistic and functional studies were performed to elucidate the role of our top candidate splice variant, BAF45d, in glioblastoma. Results: BAF45d is part of the switch/sucrose nonfermentable complex and plays a key role in the development of the CNS. The BAF45d/6A isoform is present in 85% of over 200 glioma samples that have been analyzed and contributes to the malignant glioma phenotype through the maintenance of an undifferentiated cellular state. We demonstrate that BAF45d splicing is mediated by polypyrimidine tract-binding protein 1 (PTBP1) and that BAF45d regulates PTBP1, uncovering a reciprocal interplay between RNA splicing regulation and transcription. Conclusions: Our data indicate that AS is a mechanism that contributes to the malignant phenotype of glioblastoma. Understanding the consequences of this biological process will uncover new therapeutic targets for this devastating disease.

Phase I Trial of DNX-2401 for Diffuse Intrinsic Pontine Glioma Newly Diagnosed in Pediatric Patients.

BACKGROUND: There are no effective treatments for diffuse intrinsic pontine gliomas (DIPGs); these tumors cannot be surgical resected, and diagnosis is based on magnetic resonance imaging. As a result, tumor tissues for molecular studies and pathologic diagnosis are infrequent. New clinical trials are investigating novel medications and therapeutic techniques in an effort to improve treatment of patients with DIPGs. OBJECTIVE: To determine the safety, tolerability, and toxicity of an oncolytic adenovirus, DNX-2401, injected into the cerebellar peduncle in pediatric subjects with DIPG and to collect tumor samples of this type of tumor. METHODS: Phase I, single-center, uncontrolled trial. A tumor biopsy will be performed through the cerebellar peduncle, and DNX-2401 will be injected immediately after the biopsy. Standard therapy consisting of radiotherapy and chemotherapy will follow in 2 to 6 wk. EXPECTED OUTCOMES: Improvement of overall survival and quality of life in patients with DIPG and collection of tumor specimens to study the molecular profiling of these tumors. DISCUSSION: The aims of this trial are to contribute to the sample collection of DIPG and to offer treatment during the tumor tissue biopsy using the virus. If this virus works as expected, it could kill the tumor cells with no damage to healthy tissue, functioning as a targeted therapy. It is important to note that edema has not been observed with this virus in all trials performed to date. The information obtained through this and other similar studies may be useful for developing or improving new therapies in the battle against DIPG.

A phase II trial of autologous dendritic cell vaccination and radiochemotherapy following fluorescence-guided surgery in newly diagnosed glioblastoma patients.

BACKGROUND: Prognosis of patients with glioblastoma multiforme (GBM) remains dismal, with median overall survival (OS) of about 15 months. It is therefore crucial to search alternative strategies that improve these results obtained with conventional treatments. In this context, immunotherapy seems to be a promising therapeutic option. We hypothesized that the addition of tumor lysate-pulsed autologous dendritic cells (DCs) vaccination to maximal safe resection followed by radiotherapy and concomitant and adjuvant temozolomide could improve patients' survival. METHODS: We conducted a phase-II clinical trial of autologous DCs vaccination in patients with newly diagnosed patients GBM who were candidates to complete or near complete resection. Candidates were finally included if residual tumor volume was lower than 1 cc on postoperative radiological examination. Autologous DCs were generated from peripheral blood monocytes and pulsed with autologous whole tumor lysate. The vaccination calendar started before radiotherapy and was continued during adjuvant chemotherapy. Progression free survival (PFS) and OS were analyzed with the Kaplan-Meier method. Immune response were assessed in blood samples obtained before each vaccines. RESULTS: Thirty-two consecutive patients were screened, one of which was a screening failure due to insufficient resection. Median age was 61 years (range 42-70). Karnofsky performance score (KPS) was 90-100 in 29%, 80 in 35.5% and 60-70 in 35.5% of cases. MGMT (O6-methylguanine-DNA-methyltransferase) promoter was methylated in 45.2% of patients. No severe adverse effects related to immunotherapy were registered. Median PFS was 12.7 months (CI 95% 7-16) and median OS was 23.4 months (95% CI 16-33.1). Increase in post-vaccination tumor specific immune response after vaccines (proliferation or cytokine production) was detected in 11/27 evaluated patients. No correlation between immune response and survival was found. CONCLUSIONS: Our results suggest that the addition of tumor lysate-pulsed autologous DCs vaccination to tumor resection and combined radio-chemotherapy is feasible and safe. A multicenter randomized clinical trial is warranted to evaluate the potential survival benefit of this therapeutic approach. Trial registration This phase-II trial was registered as EudraCT: 2009-009879-35 and ClinicalTrials.gov Identifier: NCT01006044 retrospectively registered.

Correlación de la capacidad antioxidante total entre suero, saliva y orina empleando las técnicas de ABTS y FRAP en personas saludables / Correlation of the total antioxidant capacity between serum, saliva and urine using the ABTS techniques and FRAP in healthy people

Introducción: La evaluación de la capacidad antioxidante total (CAT) constituye un indicador más del estado de salud. La muestra de elección es suero o plasma, su obtención es invasiva, la complejidad varía de acuerdo al tipo de paciente. Alternativamente pueden emplearse otros fluidos de obtención no invasiva como la saliva y la orina. Se requiere validar la correlación de la CAT entre estos tres tipos de fluidos. Objetivo: Establecer la correlación de la CAT entre muestras de suero, saliva y orina empleando las técnicas de ABTS+ y FRAP en personas saludables. Diseño: Observacional-analítico. Lugar: Centro de Investigación de Bioquímica y Nutrición. Facultad de Medicina, UNMSM, Perú. Participantes: Veintitrés personas voluntarias aportaron muestras de suero, saliva y orina. Intervenciones: Las muestras se almacenaron a -20 grados Centígrados. Se emplearon las técnicas de ABTS+ y de FRAP. Se obtuvo el consentimiento informado y se contó con la aprobación del Comité de Ética de la Facultad de Medicina-UNMSM. Resultados: Con la técnica de ABTS+, la correlación de Pearson para suero-saliva fue -0,315; suero-orina, 0,165 y saliva-orina, -0,150. Con la técnica del FRAP, el coeficiente de correlación de Pearson entre suero-saliva fue -0,015, suero-orina 0,206, y saliva-orina -0.086. Conclusión: No existe correlación significativa entre estos tres tipos de muestras. Sin embargo, se observa que la correlación entre suero-saliva por ABTS+ y suero-orina por FRAP son no despreciables y es probable que aumente con mayor número de voluntarios. La muestra con mayor CAT es la orina, y la de menor CAT, la saliva.

Abstract: Evaluation of total antioxidant capacity (TAC) is another indicator of health status. The specimen of choice is serum or plasma, their collection is invasive, complexity varies according to the kind of patient. Alternatively other non-invasively obtaining fluids as saliva and urine may be used. It is necessary to validate the correlation of TAC between these three types of fluids. Objective: To establish the correlation between the TAC serum, saliva and urine using the ABTS+ and FRAP techniques in health people. Design: Observational-analytic. Setting: Biochemistry and Nutrition Research. Faculty of Medicine, UNMSM, Peru. Participants: Twenty three volunteers provided samples of serum, saliva and urine. Interventions: The samples were stored at -20 degrees Celsius. ABTS+ and FRAP techniques were used. It was informed and was approved by the Ethics Committee of the Faculty of Medicine-San Marcos. The consent was obtained. Results: By the ABTS+ technique, the Pearson correlation to serum-saliva was -0,315; serum-urine, 0,165 and saliva- urine, -0,150. By FRAP technique, the Pearson correlation to serum-saliva was -0,015; serum-urine, 0,206 and saliva-urine -0,086. Conclusion: There is not significant correlation between these three types of samples. However, we can mention that the correlation between serum-saliva by ABTS+ and serum-urine by FRAP are not negligible and is likely to increase with more volunteers. The sample with greatest TAC is the urine, and the lowest TAC, the saliva.

A small noncoding RNA signature found in exosomes of GBM patient serum as a diagnostic tool.

BACKGROUND: Glioblastoma multiforme (GBM) is the most frequent malignant brain tumor in adults, and its prognosis remains dismal despite intensive research and therapeutic advances. Diagnostic biomarkers would be clinically meaningful to allow for early detection of the tumor and for those cases in which surgery is contraindicated or biopsy results are inconclusive. Recent findings show that GBM cells release microvesicles that contain a select subset of cellular proteins and RNA. The aim of this hypothesis-generating study was to assess the diagnostic potential of miRNAs found in microvesicles isolated from the serum of GBM patients. METHODS: To control disease heterogeneity, we used patients with newly diagnosed GBM. In the discovery stage, PCR-based TaqMan Low Density Arrays followed by individual quantitative reverse transcriptase polymerase chain reaction were used to test the differences in the miRNA expression levels of serum microvesicles among 25 GBM patients and healthy controls paired by age and sex. The detected noncoding RNAs were then validated in another 50 GBM patients. RESULTS: We found that the expression levels of 1 small noncoding RNA (RNU6-1) and 2 microRNAs (miR-320 and miR-574-3p) were significantly associated with a GBM diagnosis. In addition, RNU6-1 was consistently an independent predictor of a GBM diagnosis. CONCLUSIONS: Altogether our results uncovered a small noncoding RNA signature in microvesicles isolated from GBM patient serum that could be used as a fast and reliable differential diagnostic biomarker.