Glioblastoma (GBM) is the most common and aggressive primary brain tumor and is comprised of transcriptionally heterogeneous cells and a complex microenvironment. Despite decades of research effort, few treatments significantly benefit clinical outcomes. This may be, in part, due to the lack of tools to directly measure functional responses of these heterogeneous cell types under therapy. This thesis aims to advance the understanding of cell type-specific therapeutic response by the development and application of integrated single cell imaging and RNA sequencing technology. Chapter 1 provides an overview of GBM and its heterogeneity, how investigation of cell type-specific phenotypes would benefit the development of GBM treatments, and current sequencing and imaging technologies to examine cell phenotypes with single-cell resolution. Chapter 2 presents a new microfluidic technology for joint single cell imaging and RNA sequencing that can link imaging-based phenotypes and transcriptional identity of the same individual cells with high throughput, molecular capture efficiency, linking accuracy, and user-friendliness. Chapters 3 and 4 present applications of this technology in investigating cell type-specificities of GBM treatments. Chapter 3 focuses on the specificities of 5-aminolevulinic acid (5-ALA), an FDA approved fluorogenic agent, used in fluorescence guided surgery and reveals 5-ALA labeling is not specific to transformed glioma cells, which encourages further studies to systematically compare its performance with potential alternatives. Chapter 4 focuses on the specificities of drug responses by presenting a functional drug screening approach that directly links cell states measured by apoptosis indicators with transcriptional states, which greatly enhances the interpretability of single cell-resolved drug perturbation assays.

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.

Bakgrund Under 2021 drabbades ungefär 1400 personer i Sverige av en tumör i det centrala nervsystemet, vilket ledde till 675 dödsfall. Hjärntumörer kan orsaka svåra symtom som huvudvärk, aptitförlust, personlighetsförändringar och psykiska problem. Sjuksköterskan ansvarar då för att hjälpa patienten men också att stödja de anhöriga som går igenom en utmanande tid. I och med att patientens självständighet minskar, hamnar anhöriga ofta i en ny roll med ökat ansvar. Detta understryker behovet av att belysa anhörigas upplevelser och stödja dem på ett värdigt sätt. Syfte Syftet med studien var att belysa anhörigvårdarens upplevelser av att vårda en patient med hjärntumör. Metod En icke-systematisk litteraturöversikt baserad på 10 vetenskapliga artiklar. Sökningarna gjordes på PubMed och CINAHL. De valda artiklarna granskades Sophiahemmet högskolas bedömningsmall. En integrerad analys användes för att analysera och sammanställa resultatet. Resultat Fyra kategorier framkom efter analysen av materialet. Dessa var vårdgivarens börda, känslomässiga reaktioner, relationer samt stöd. Resultatet visade att anhöriga hade mycket negativa upplevelser från att ha vårdat sina anhöriga med hjärntumörer. Anhörigvårdare upplevde omfattande förändringar i vardagen, där de fick andra ansvar som de inte haft tidigare vilka krävde tid, energi och tillgänglighet. Denna förändring medförde starka känslomässiga reaktioner, förändringar i relationer och ett stort stödbehov. Slutsats De anhörigas upplevelser var främst negativa och de skulle behöva mer stöd och hjälp för att klara av den befintliga situationen bättre. Litteraturöversikten identifierade att de anhöriga upplevde starka känslomässiga reaktioner, förändringar i relationer, en börda i det ansvar de behöver ta och ett behov av mer stöd. Med denna ökade kunskap och förståelse kan vårdpersonalen hjälpa till och förhoppningsvis också lyckas minska de negativa konsekvenserna. / Background During 2021, approximately 1400 people in Sweden were diagnosed with central nervous system tumors, resulting in 675 deaths. Brain tumors can cause severe symptoms such as headaches, decreased appetite, personality changes and psychological problems. Nurses are responsible to help the patient and supporting the family during this challenging time. As the patient's independence diminishes, relatives often find themselves in a new role with increased responsibilities. This highlights the need to illuminate the experiences of family members and support them in a dignified manner. Aim This study aims to explore family caregivers' experiences of taking care of a patient with a brain tumor. Method A non-systematic review of 10 studies was conducted using PubMed and CINAHL databases. The selected articles were reviewed using an assessment template. An integrated analysis was used to analyze and compile the results. Results Four categories emerged after the analysis of the material. These were caregiver burden, emotional reactions, relationships, and support. The results showed that relatives had very negative experiences from caring for their relatives with brain tumors. Family caregivers experienced extensive changes in their everyday lives, where they were given other responsibilities they had not previously had, requiring time, energy, and availability. This change brought strong emotional reactions, changes in relationships, and a great need for support. Conclusions The family's experiences were mainly negative and they would need more support and help to be able to cope with the situation they have been put in in a better way. The literature review identified that the family experienced strong emotional reactions, changes in relationships, a burden in new responsibilities they got and a need for more support. With this increased knowledge and understanding, the healthcare staff can help and hopefully also succeed in reducing the negative consequences.

2023 / Connectome Fingerprinting (CF) and Connectome-Based Predictive Modeling (CPM) are emerging data science approaches within cognitive neuroscience that utilize brain connectivity data to predict the functional organization of the brain and behavior. Here, I examined the robustness and data needs of these approaches and developed applications for specific clinical populations, including neurosurgical glioma patients and early-stage Alzheimer’s Disease (AD) patients.In the first study, I utilized the Human Connectome Project dataset (n=208) to understand the relationship between function and connectivity using CF across cognitive tasks, and functional connectivity paradigms of fixation and movie-watching. I benchmarked the CF method to establish the quantity and quality of connectivity and task activation data required to build robust models across different cortical regions, cognitive tasks, scan quality, learning algorithms, and scanners. I also modeled the link of cerebellar-cortical connectivity with function, suggesting an integrated role of the cerebellum across cognitive domains. Neurosurgeons routinely use task fMRI protocols to map out motor and language networks to aid in presurgical planning for brain tumor resection to minimize cognitive damage. In the second study, I investigated the possibility of using CF models to help neurosurgeons predict motor and language networks in presurgical patients with gliomas (n=16), who are unable to perform complex tasks in the scanner. I tested the model validity across healthy control adults (n=16) from different scanners with varying data quality and quantity. The deposition of tau and amyloid-β plaques in AD results in brain degeneration, memory loss, and behavioral changes. Resting-state functional connectivity gets affected due to the progression of AD. In the third study, I analyzed the relationship between functional connectivity and tau/amyloid depositions and tested the predictive ability of CPM across two different cohorts: Autosomal Dominant with Presenilin1 mutation from the Colombia-Boston study with early-onset AD (n=32) and late-onset sporadic AD with APOE4 marker from the Harvard Aging Brain Study (n=78) and compared them with healthy age-matched adults (n=35/n=206) and healthy young adults (n=1570) from the Genomic Superstruct Project. Combined, these studies highlight the potential of data-driven approaches to model human brain function and connectivity, facilitating fMRI's translational applications. / 2027-02-04T00:00:00Z

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.