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No description available.

Histotripsy is a non-thermal, non-invasive focused ultrasound therapy using controlled acoustic cavitation to mechanically disintegrate tissue into an acellular homogenate. Histotripsy applies microsecond-length, high pressure (> 10 MPa) pulses to initiate the rapid expansion and collapse of nuclei in a millimeter-scale focal region, applying large stresses and strains to targeted tissues. The cavitation "bubble cloud" generated during histotripsy treatment can be visualized in real time on ultrasound imaging, assisting with treatment guidance and monitoring. Past studies have demonstrated histotripsy's potential for a variety of applications, but histotripsy has not yet been investigated for superficial musculoskeletal tumor ablation. Additionally, preliminary investigations using histotripsy to ablate brain tumors are underway, but require advanced histotripsy devices capable of overcoming attenuation of the therapeutic ultrasound signal by the skull and rely on MRI for real-time guidance. As a result, open questions remain regarding ultrasound-guided histotripsy for brain tumors. Early evidence also suggests that histotripsy ablation may induce immunogenic changes in the tumor microenvironment. Continued research is needed to explain and corroborate these findings under conditions more immunologically representative of human cancers, such as in large animal models with spontaneous tumors. This dissertation investigates the safety and feasibility of using ultrasound-guided histotripsy to ablate superficial soft tissue sarcomas (STS), osteosarcomas (OS), and brain tumors and considers the immunological impacts of histotripsy treatment for STS and OS. The research described herein (1) investigates the ability of histotripsy to treat superficial STS tumors in companion animals with spontaneous tumors, (2) investigates the feasibility of treating bone tumors with histotripsy through a series of ex vivo and in vivo studies, and (3) applies histotripsy for the minimally invasive treatment of superficial brain tumors. The completion of this dissertation will provide significant insight into the ability of ultrasound-guided histotripsy to treat novel tumor types (i.e., STS, OS, and brain tumors) and the potential role of histotripsy in veterinary medicine. Future work will build upon the studies detailed in this dissertation to optimize ultrasound-guided histotripsy for the treatment of complete STS, OS, and brain tumors in veterinary and human patients. / Doctor of Philosophy / Histotripsy is a non-invasive focused ultrasound therapy that mechanically breaks down targeted tissues through acoustic cavitation. Histotripsy is currently being developed for a number of clinical applications, including tumor ablation, but its potential for treating many cancer types remains unknown. Histotripsy uses very short, high pressure ultrasound pulses to initiate the nucleation of bubbles in the target region. These bubbles then expand and rapidly collapse to impart large stresses and strains on surrounding tissues, leaving behind only acellular debris. The cavitation "bubble cloud" generated during histotripsy treatment can be visualized on ultrasound imaging, offering real-time treatment guidance and monitoring. Histotripsy has not yet been investigated for superficial musculoskeletal tumor ablation, and preliminary studies using histotripsy to ablate brain tumors are underway, but require advanced histotripsy devices still under development. As a result, open questions remain regarding histotripsy ablation as a treatment for musculoskeletal and brain tumors. Additionally, early evidence suggests that histotripsy ablation may be able to stimulate an immune response, treating not only the targeted tumor but also multifocal or metastatic disease. Continued research is needed to explain and corroborate these findings under conditions more similar to human cancers, such as in large animal models with naturally-occurring tumors. This dissertation investigates the safety and feasibility of using ultrasound-guided histotripsy to ablate superficial soft tissue sarcomas (STS), osteosarcomas (OS), and brain tumors and considers the immunological impacts of histotripsy treatment for STS and OS. This research (1) investigates the ability of histotripsy to treat superficial STS tumors in companion animals with spontaneous tumors, (2) investigates the feasibility of treating bone tumors with histotripsy through a series of ex vivo and in vivo studies, and (3) applies histotripsy for the minimally invasive treatment of superficial brain tumors. The completion of this dissertation will provide significant insight into the ability of ultrasound-guided histotripsy to treat novel tumor types (i.e., STS, OS, and brain tumors) and the potential role of histotripsy in veterinary medicine. Future work will build upon the studies detailed in this dissertation to optimize ultrasound-guided histotripsy for the treatment of complete STS, OS, and brain tumors in veterinary and human patients.

Functional proton-beam stereotactic radiosurgery requires sub-millimeter alignment accuracy. A patient tracking system called Sequential Alignment and Position Verification System (SAVPS) is under development at Loma Linda University Medical Center. An optical positioning system (OPS), manufactured by Vicon Peak, has been chosen to verify the correct alignment of the target with the proton beam axis. The main objective of this thesis is to optimize an existing version of SAVPS by conducting error analysis. An image processing algorithm was developed and applied to estimate the error introduced by the Patient Positioning System (PPS) in order to derive the true error of the SAVPS.

Os gliomas são tumores do sistema nervoso central (SNC) que evoluem a partir das células da glia. O tipo mais frequente e mais agressivo destes tumores é conhecido como glioblastoma multiforme (GBM) e entre as características biológicas de agressividade associadas a esse tumor estão o seu rápido crescimento e ausência de apoptose. O seu prognóstico desfavorável está associado à dificuldade de tratamento dessas células, pois possuem resistência à quimioterapia e a radioterapia. A expressão gênica das enzimas ciclooxigenase-1 (COX-1), ciclooxigenase-2 (COX-2), prostaglandina E sintase-1 microssomal (mPGES-1), prostaglandina E sintase-2 microssomal (mPGES-2), prostaglandina E sintase citosólica (cPGES) e os produtos da síntese destas enzimas, incluindo a prostaglandina E1 (PGE1) e a prostaglandina E2 (PGE2) estão diretamente relacionados com a malignidade dos gliomas. A PGE1 e a PGE2 podem atuar de modo autócrino e parácrino, interagindo com suas células alvos através de ligação aos receptores da superfície celular que estão ligados a proteína G. Estes receptores são conhecidos como receptores EPs e dividem-se em quatro subtipos: EP-1, EP-2, EP-3 e EP-4 sendo que cada um deles ativa vias distintas de sinalização intracelular. Desta forma, este estudo teve por objetivo analisar in vitro o papel da PGE1, PGE2 e seus receptores na proliferação e apoptose em glioma humano, e a expressão das enzimas COX-1, COX-2, mPGES-1, mPGES-2 e cPGES. / Gliomas are tumors of the central nervous system (CNS) that evolve from glial cells. The most common and most aggressive form of these tumors is known as glioblastoma multiforme (GBM). The biological aggressiveness of GBM is associated with its rapid growth and lack of apoptosis. Its poor prognosis is strongly associated with the difficulty of treating these cells as they are resistant to chemotherapy and radiotherapy. The gene expression of the enzymes cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), microsomal prostaglandin E synthase-1 (mPGES-1), microsomal prostaglandin E synthase-2 (mPGES-2), cytosolic prostaglandin E synthase (cPGES) and the products of the activity of these enzymes, including prostaglandin E1 (PGE1) and prostaglandin E2 (PGE2), are directly related to the malignancy of gliomas. PGE1 and PGE2 can act in an autocrine and paracrine manner, by interacting with their target cells via binding to cell surface receptors that are linked to G-proteins. These receptors are known as EP receptors and are divided into four subtypes: EP1, EP2, EP3 and EP4; each of which activates distinct intracellular signaling pathways. Therefore, this study aimed to analyze, in vitro, the role of PGE1, PGE2 and their receptors in the proliferation and apoptosis of human glioma and the expression of COX-1, COX-2, mPGES-1, mPGES-2 and cPGES.

Meduloblastoma é o tumor intracranial mais comum em crianças, provavelmente derivado de células precursoras da camada granular externa do cerebelo durante seu desenvolvimento. O tratamento padrão consiste em cirurgia, radioterapia e quimioterapia, que produzem graves sequelas nos pacientes e garantem uma sobrevida baixa, o que demonstra a necessidade de novas alternativas terapêuticas para a doença. Evidências demonstram que o receptor do peptídeo liberador de gastrina (GRPR) está superexpresso em diversos tumores humanos, assim como seu agonista (GRP) pode atuar como um fator de crescimento autócrino em tumores cerebrais. No presente estudo, avaliamos a expressão de GRPR e o efeito de seus agonistas, bombesina (BB) e GRP, além do antagonista RC-3095, sobre a viabilidade celular de linhagens de meduloblastoma humano DAOY, D283 e ONS76. Mostramos que meduloblastomas, apesar de expressarem GRPR, não têm sua viabilidade celular afetada por agonistas e antagonista desse receptor. Uma vez que há evidências de que BDNF (fator neurotrófico derivado de cérebro) esteja relacionado à diferenciação celular em meduloblastomas, também avaliamos o efeito de BDNF sobre a viabilidade celular das linhagens de meduloblastoma humano. As linhagens DAOY e D283 tiveram sua viabilidade celular reduzida pela presença de BDNF. Uma vez que a via da PKA tem sido implicada na iniciação e progressão de vários tumores, também avaliamos o efeito de rolipram, um inibidor de fosfodiesterase tipo IV, sobre a viabilidade celular das linhagens de meduloblastoma humano, sendo que rolipram reduziu a viabilidade celular de todas as linhagens estudadas. Os receptores de BDNF e a via da PKA podem, portanto, ser alvos moleculares promissores para o desenvolvimento de novas terapias para meduloblastomas. / Medulloblastoma is the most common intracranial tumor in children and is believed to arise from the precursor cells of the external granule layer of the developing cerebellum. The standard treatment, consisting of surgery, craniospinal radiotherapy and chemotherapy, produces severe sequelae in patients and provides a poor overall survival, indicating the need for new therapeutic alternatives for treating this disease. Evidences show that the gastrin releasing peptide receptor (GRPR) is overexpressed in various human tumors and its agonist (GRP) can act as an autocrine growth factor in brain tumors. In the present study, we evaluated GRPR expression, as well as the effect of its agonists, bombesin (BB) and GRP, and its antagonist RC-3095, over cell viability of the human medulloblastoma cell lines DAOY, D283 and ONS76. We found that medulloblastomas, in spite of expressing GRPR, do not have its viability affected by the presence of agonists and antagonist of this receptor. Since there are evidences that BDNF (brain-derived neurotrophic factor) is related to cell differentiation in medulloblastomas, we also evaluated the effect of BDNF over the viability of medulloblastoma cell lines. The viability of the cell lines DAOY and D283 was reduced by the presence of BDNF. Since the PKA pathway has been implicated in the initiation and progression of various tumors, we also evaluated the effect of rolipram, a phosphodiesterase IV inhibitor, over the viability of the same medulloblastoma cell lines and we found that rolipram inhibited the viability of all the cell lines studied. BDNF receptors, as well as the PKA pathway, may be therefore promising molecular targets for the development of new therapies for treating medulloblastomas.

Glioblastoma multiforme (GBM) is a debilitating malignant brain tumor with expected patient survival of less than a year and limited responsiveness to most treatments, often requiring biopsy for diagnosis and invasive surgery for treatment. We propose a DNA computer system, consisting of input, computation, and output components, for diagnosis and treatment. The input component will detect the presence of three GBM biomarkers: vascular endothelial growth factor (VEGF), caveolin-1α (CAV), and B2 receptors. The computation component will include indicator segments for each of these genes, and ensure that output is only released if all the biomarkers are present. The output component will consist of the therapeutic agent interleukin-12 (IL-12). This study will designate four groups of animals: untreated tumor-free (control), tumor-inoculated (RG2), treated and tumor-free (DNA), and treated and tumor-inoculated (RG2/DNA). In the RG2 and RG2/DNA groups, we will inoculate adult male Fischer rats with RG2 cells into the striatum to induce tumor growth. Rats in the DNA and RG2/DNA groups will be implanted with the DNA system at the same location via recombinant adeno- associated viral vectors. The effectiveness of the DNA system will be evaluated through tumor size measurements, collected from brain slices stained with hematoxylin and eosin, and survival curve. Additionally, IL-12 localization will confirm the release of the output component. We anticipate that the DNA treatment will result in a decrease in tumor size, leading to smaller tumor size in the RG2/DNA group versus the RG2 group. The control group is expected to survive the longest, followed by the DNA group, then the RG2/DNA group, and finally the RG2 group. In the DNA group, IL-12 is expected to stay localized to the implantation site, remaining in its unreleased stem-loop form. On the other hand, it is expected to be released and active in the RG2/DNA group. This study provides a proof of concept to demonstrate the viability and effectiveness of a DNA system using VEGF, CAV, and B2 receptors as biomarkers and IL-12 as a therapeutic output component in the RG2 model. Further research may include varying several of the parameters used in this study, including amount of RG2 administered, choice of biomarkers, quantity and choice of output component, and choice of animal model. This system provides a promising and innovative new approach that is less invasive than surgery yet is still effective in diagnosing, targeting, and treating GBM.

Η εισαγωγή της Μαγνητικής Τομογραφίας (ΜΤ) στην κλινική πρακτική και η συμπληρωματική πληροφορία που δίνει η Φασματοσκοπία Μαγνητικού Συντονισμού (ΦΜΣ) συνιστά μια από τις πιο σημαντικές εξελίξεις στη διάγνωση ασθενών με καρκίνο εγκεφάλου [1]. Παρ’ όλα αυτά, οι εικόνες ΜΤ είναι συχνά δύσκολο να ερμηνευθούν από τους ειδικούς λόγω [2] α/ της υποκειμενικότητας και περιορισμένης εμπειρίας του παρατηρητή στην εκτίμηση εικόνων που παράγει η σχετικά νέα αυτή τεχνολογία, β/ των ποικίλων κλινικών χαρακτηριστικών των όγκων (π.χ. τύπος, διαβάθμιση κακοήθειας κλπ.) και γ/ της ιδιαιτερότητας των όγκων στην αντίθεση που παρουσιάζουν με τον περιβάλλοντα ιστό. Μόνο λιγοστές μελέτες έχουν διεξαχθεί για να χαρακτηρίσουν ιστούς εγκεφάλου μέσω της ανάλυσης ποσοτικών χαρακτηριστικών από εικόνες εγκεφάλου ΜΤ [3, 4]. Ενώ έχει ήδη τονιστεί η αναγκαιότητα συσχετισμού της διαγνωστικής και προγνωστικής πληροφορίας που προέρχεται από εικόνες ΜΤ και σήματα ΦΜΣ στη διεθνή βιβλιογραφία [5], υπάρχουν λιγοστές ανάλογες αναφορές για τον σχεδιασμό και υλοποίηση συστήματος Η/Υ αυτόματης διάγνωσης όγκων εγκεφάλου κάνοντας συνδυασμό ποσοτικής πληροφορίας προερχόμενης από εικόνες ΜΤ και σήματα ΦΜΣ [6, 7]. Οι στόχοι της παρούσας διατριβής εστιάζονται στα παρακάτω: - στη μελέτη, ανάπτυξη και η υλοποίηση υπολογιστικού συστήματος αυτόματης ταξινόμησης όγκων του εγκεφάλου μέσω της ποσοτικής ανάλυσης εικόνων ΜΤ το οποίο θα βελτιώνει την ακρίβεια ταξινόμησης σε σχέση με ήδη υπάρχοντα συστήματα [4, 8, 9], όπως αυτά περιγράφονται στην διεθνή βιβλιογραφία μεταξύ πρωτογενών και δευτερογενών όγκων εγκεφάλου καθώς και μεταξύ γλοιωμάτων και μηνιγγιωμάτων με την χρήση δέντρου ιεραρχικής απόφασης δύο επιπέδων. Επιπλέον, στην ανάδειξη πως η χρήση ενός μη-γραμμικού πολυωνυμικού μετασχηματισμού ελάχιστων τετραγώνων των χαρακτηριστικών υφής έχει ως αποτέλεσμα την βελτίωση της ακρίβειας ταξινόμησης του ταξινομητή πιθανοκρατικού νευρωνικού δικτύου. - στην επέκταση και την βελτίωση του συστήματος αυτόματης ταξινόμησης όγκων του εγκεφάλου χρησιμοποιώντας α/ ογκομετρικές ποσοτικές παραμέτρους εικόνων ΜΤ, β/ ταξινομητή μηχανών διανυσμάτων στήριξης μαζί με τη μεθοδολογία συνάθροισης αποτελεσμάτων ταξινόμησης από τυχαιοποιημένα δείγματα κατηγοριών δημιουργημένων με επαναδειγματοληψία για κάθε κόμβο δέντρου ιεραρχικής απόφασης δύο επιπέδων όπου στο πρώτο επίπεδο πραγματοποιήθηκε διαχωρισμός μεταξύ πρωτογενών και δευτερογενών όγκων εγκεφάλου και στο δεύτερο και μεταξύ γλοιωμάτων και μηνιγγιωμάτων και γ/ έναν τροποποιημένο πυρήνα ακτινικής συνάρτησης βάσης για τον ταξινομητή μηχανών διανυσμάτων στήριξης ο οποίος περιλαμβάνει την τεχνική μη-γραμμικού πολυωνυμικού μετασχηματισμού ελάχιστων τετραγώνων με στόχο την βελτίωση της ακρίβειας ταξινόμησης. - στην περαιτέρω επέκταση και την βελτίωση του συστήματος αυτόματης ταξινόμησης με την εισαγωγή χαρακτηριστικών προερχόμενων από σήματα ΦΜΣ ώστε να διερευνηθεί εάν η χρήση του μπορεί να βελτιώσει τα αποτελέσματα ταξινόμησης μεταξύ μηνιγγιωμάτων και μονήρων μεταστάσεων. Τέλος κάνοντας μια περίληψη, η παρούσα διατριβή διαπραγματεύεται τον σχεδιασμό, ανάπτυξη και υλοποίηση μεθόδων και αλγορίθμων για την επεξεργασία και ανάλυση ιατρικών εικόνων, επικεντρώνοντας ειδικότερα στην εφαρμογή των μεθόδων αυτών για την διάγνωση του τύπου των όγκων εγκεφάλου. Τα πιο βασικά συμπεράσματα που απορρέουν από την παρούσα διατριβή είναι τα ακόλουθα: α/ Το σύστημα ταξινόμησης των τύπων των όγκων εγκεφάλου που σχεδιάστηκε και υλοποιήθηκε αυξάνει τα ποσοστά ορθής ταξινόμησης σε σχέση με τα υπάρχοντα. β/ Η κωδικοποίηση των ιδιοτήτων της υφής που προέρχεται από τον σύνολο του όγκου παρέχει επιπρόσθετη πληροφορία στο σύστημα ταξινόμησης αυξάνοντας τα ποσοστά επιτυχούς διαχωρισμού. γ/ Τα χαρακτηριστικά φασματοσκοπίας μαγνητικού συντονισμού αποτελούν επιπρόσθετη αξία στο χαρακτηρισμό του τύπου των όγκων εγκεφάλου μιας και οδήγησαν στην αύξηση του ποσοστού επιτυχούς διαχωρισμού του συστήματος ταξινόμησης. / The process of brain tumor characterization requires a rather intricate assessment of the various Magnetic Resonance (MR) image and spectroscopic features and is typically performed by experienced radiologists. Despite the inherently subjective nature of many of the decisions associated with this process, an expert radiologist is able to perform this task with a significant degree of precision and accuracy. However, in the effort to deliver more effective treatment, clinicians are continuously seeking for greater accuracy in the pathological characterization of brain tissues. The aim of the present thesis was to design, implement, and evaluate a software classification system for discriminating between different brain tumor types on Magnetic Resonance Imaging (MRI), employing textural and spectroscopic features. The clinical material consisted of sixty seven T1-weighted post-contrast MR brain images (21 metastases, 19 meningiomas, and 27 gliomas), obtained from patients with verified and untreated intracranial tumors. Thirty-six 2-dimensional textural features (2D), from the image histogram and the co-occurrence and run-length matrices, were extracted from each one of 67 MR-images. Similarly, an equal number of 3-dimensional textural features (3D) were also calculated in the attempt to maximize classification performances. Finally, MR-spectroscopy features were also incorporated for improving classification accuracies. Classification methods employed included i/ a modified Probabilistic Neural Network (PNN) and Support Vector Machines (SVM) algorithms, incorporating a non-linear Least Squares Features Transformation (LSFT) into the classifiers and ii/ an ensemble classification scheme employing the LSFT-SVM classifier. The LSFT improved classifiers’ performances, increased class separability, and resulted in dimensionality reduction. For evaluating the performance of the designed classification schemes, evaluations were performed by means of the external cross validation process, which is considered indicative of the generalization performance of the designed classification system to ‘unseen’ cases. It was found that the LSFT features transformation enhanced the performance of the PNN and SVM algorithms, achieving classification accuracies of 73.48 % in distinguishing metastatic from primary tumors and 88.67% in discriminating gliomas from meningiomas. When volumetric 3-dimensional features were employed, these results improved to 88.18% for discriminating between metastatic and primary tumors and 97.33% for distinguishing gliomas from meningiomas. The textural features employed in the design of the optimum classification scheme were associated primarily with image texture homogeneity. Finally, when MR-spectroscopy features were also incorporated, classification accuracy was boosted up from 95% in discriminating meningiomas from metastasis to 100%. The MR-image features that participated in the optimum feature vector were related to the degree of homogeneity, the amount of randomness and the dispersion of the gray-tone intensity values within the texture of the tumor. These textural characteristics are related to textural parameters that physicians employ in diagnosis and they were proportional to the textural imprint of brain tumors, i.e. gliomas have heterogeneous texture while meningiomas appear to be homogeneous in MR imaging. The MR-spectroscopy feature that participated in the optimum feature vector was the Choline (Cho) / N-Acetyl Aspartate (NAA) metabolite integral ratio. It was found that both meningiomas and metastases are characterized from low concentrations of NAA while meningiomas exhibit higher concentrations of Cho than metastases, which could be attributed to increased synthesis of tumor cell membranes. Finally, the proposed system might be of value as an assisting tool for brain tumor characterization on volumetric MRI series.

Meduloblastoma é o tumor intracranial mais comum em crianças, provavelmente derivado de células precursoras da camada granular externa do cerebelo durante seu desenvolvimento. O tratamento padrão consiste em cirurgia, radioterapia e quimioterapia, que produzem graves sequelas nos pacientes e garantem uma sobrevida baixa, o que demonstra a necessidade de novas alternativas terapêuticas para a doença. Evidências demonstram que o receptor do peptídeo liberador de gastrina (GRPR) está superexpresso em diversos tumores humanos, assim como seu agonista (GRP) pode atuar como um fator de crescimento autócrino em tumores cerebrais. No presente estudo, avaliamos a expressão de GRPR e o efeito de seus agonistas, bombesina (BB) e GRP, além do antagonista RC-3095, sobre a viabilidade celular de linhagens de meduloblastoma humano DAOY, D283 e ONS76. Mostramos que meduloblastomas, apesar de expressarem GRPR, não têm sua viabilidade celular afetada por agonistas e antagonista desse receptor. Uma vez que há evidências de que BDNF (fator neurotrófico derivado de cérebro) esteja relacionado à diferenciação celular em meduloblastomas, também avaliamos o efeito de BDNF sobre a viabilidade celular das linhagens de meduloblastoma humano. As linhagens DAOY e D283 tiveram sua viabilidade celular reduzida pela presença de BDNF. Uma vez que a via da PKA tem sido implicada na iniciação e progressão de vários tumores, também avaliamos o efeito de rolipram, um inibidor de fosfodiesterase tipo IV, sobre a viabilidade celular das linhagens de meduloblastoma humano, sendo que rolipram reduziu a viabilidade celular de todas as linhagens estudadas. Os receptores de BDNF e a via da PKA podem, portanto, ser alvos moleculares promissores para o desenvolvimento de novas terapias para meduloblastomas. / Medulloblastoma is the most common intracranial tumor in children and is believed to arise from the precursor cells of the external granule layer of the developing cerebellum. The standard treatment, consisting of surgery, craniospinal radiotherapy and chemotherapy, produces severe sequelae in patients and provides a poor overall survival, indicating the need for new therapeutic alternatives for treating this disease. Evidences show that the gastrin releasing peptide receptor (GRPR) is overexpressed in various human tumors and its agonist (GRP) can act as an autocrine growth factor in brain tumors. In the present study, we evaluated GRPR expression, as well as the effect of its agonists, bombesin (BB) and GRP, and its antagonist RC-3095, over cell viability of the human medulloblastoma cell lines DAOY, D283 and ONS76. We found that medulloblastomas, in spite of expressing GRPR, do not have its viability affected by the presence of agonists and antagonist of this receptor. Since there are evidences that BDNF (brain-derived neurotrophic factor) is related to cell differentiation in medulloblastomas, we also evaluated the effect of BDNF over the viability of medulloblastoma cell lines. The viability of the cell lines DAOY and D283 was reduced by the presence of BDNF. Since the PKA pathway has been implicated in the initiation and progression of various tumors, we also evaluated the effect of rolipram, a phosphodiesterase IV inhibitor, over the viability of the same medulloblastoma cell lines and we found that rolipram inhibited the viability of all the cell lines studied. BDNF receptors, as well as the PKA pathway, may be therefore promising molecular targets for the development of new therapies for treating medulloblastomas.

Meduloblastoma é o tumor intracranial mais comum em crianças, provavelmente derivado de células precursoras da camada granular externa do cerebelo durante seu desenvolvimento. O tratamento padrão consiste em cirurgia, radioterapia e quimioterapia, que produzem graves sequelas nos pacientes e garantem uma sobrevida baixa, o que demonstra a necessidade de novas alternativas terapêuticas para a doença. Evidências demonstram que o receptor do peptídeo liberador de gastrina (GRPR) está superexpresso em diversos tumores humanos, assim como seu agonista (GRP) pode atuar como um fator de crescimento autócrino em tumores cerebrais. No presente estudo, avaliamos a expressão de GRPR e o efeito de seus agonistas, bombesina (BB) e GRP, além do antagonista RC-3095, sobre a viabilidade celular de linhagens de meduloblastoma humano DAOY, D283 e ONS76. Mostramos que meduloblastomas, apesar de expressarem GRPR, não têm sua viabilidade celular afetada por agonistas e antagonista desse receptor. Uma vez que há evidências de que BDNF (fator neurotrófico derivado de cérebro) esteja relacionado à diferenciação celular em meduloblastomas, também avaliamos o efeito de BDNF sobre a viabilidade celular das linhagens de meduloblastoma humano. As linhagens DAOY e D283 tiveram sua viabilidade celular reduzida pela presença de BDNF. Uma vez que a via da PKA tem sido implicada na iniciação e progressão de vários tumores, também avaliamos o efeito de rolipram, um inibidor de fosfodiesterase tipo IV, sobre a viabilidade celular das linhagens de meduloblastoma humano, sendo que rolipram reduziu a viabilidade celular de todas as linhagens estudadas. Os receptores de BDNF e a via da PKA podem, portanto, ser alvos moleculares promissores para o desenvolvimento de novas terapias para meduloblastomas. / Medulloblastoma is the most common intracranial tumor in children and is believed to arise from the precursor cells of the external granule layer of the developing cerebellum. The standard treatment, consisting of surgery, craniospinal radiotherapy and chemotherapy, produces severe sequelae in patients and provides a poor overall survival, indicating the need for new therapeutic alternatives for treating this disease. Evidences show that the gastrin releasing peptide receptor (GRPR) is overexpressed in various human tumors and its agonist (GRP) can act as an autocrine growth factor in brain tumors. In the present study, we evaluated GRPR expression, as well as the effect of its agonists, bombesin (BB) and GRP, and its antagonist RC-3095, over cell viability of the human medulloblastoma cell lines DAOY, D283 and ONS76. We found that medulloblastomas, in spite of expressing GRPR, do not have its viability affected by the presence of agonists and antagonist of this receptor. Since there are evidences that BDNF (brain-derived neurotrophic factor) is related to cell differentiation in medulloblastomas, we also evaluated the effect of BDNF over the viability of medulloblastoma cell lines. The viability of the cell lines DAOY and D283 was reduced by the presence of BDNF. Since the PKA pathway has been implicated in the initiation and progression of various tumors, we also evaluated the effect of rolipram, a phosphodiesterase IV inhibitor, over the viability of the same medulloblastoma cell lines and we found that rolipram inhibited the viability of all the cell lines studied. BDNF receptors, as well as the PKA pathway, may be therefore promising molecular targets for the development of new therapies for treating medulloblastomas.