Biology subjects

Cruz, T.

Publications and source records attributed to Cruz, T..

4 recordsLinked to original sources

bioRxiv2026

Focused Ultrasound Blood-Tumor Barrier Opening Rapidly Augments Intratumor CD4 and CD8 T Cell Representation in a Genetically Engineered Mouse Model of Glioma

Glioblastoma (GBM) is a highly aggressive primary brain tumor that remains difficult to treat due in part to its disorganized and heterogeneous vasculature, known as the blood-tumor barrier (BTB), which limits therapeutic delivery and beneficial immune cell infiltration. Focused ultrasound (FUS) with microbubbles (MBs) can transiently disrupt the BTB to enhance drug delivery and may induce sterile inflammation (SI) that can beneficially remodel the tumor immune landscape. However, this concept has only been explored in implanted tumor models with modest immune effects. Here, we utilized a physiologically relevant genetically engineered mouse model (GEMM) generated via in utero electroporation targeting Nf1, Tp53, and Pten to study tumor-vascular-immune interactions. This 3x CRISPR-Cas9 GEMM recapitulates key features of human glioma, including infiltrative growth, histopathology, molecular alterations, and stage-dependent blood-brain barrier disruption. FUS+MBs were applied to transiently disrupt the BTB, and MRI confirmed increased vascular permeability in treated tumors. Flow cytometry revealed robust increases in tumor-infiltrating CD4+ helper and CD8+ effector T cells three days post-FUS treatment, without altering the CD8/Treg ratio. These findings were supported by immunofluorescence imaging. Double-negative and double-positive T cells were detected, but they were not significantly altered by FUS. Ki67 analysis indicated that increased T-cell accumulation was not driven by local proliferation. By seven days post-treatment, immune differences were no longer observed. Collectively, these results demonstrate that FUS-mediated BTB disruption selectively and rapidly enhances lymphocyte infiltration in a clinically relevant glioma model, supporting its potential as a temporally controlled immunomodulatory strategy for GBM.

cancer biology↗

bioRxiv2024

Focused Ultrasound Blood-Brain Barrier Opening Arrests the Growth and Formation of Cerebral Cavernous Malformations

BACKGROUNDCerebral cavernous malformations (CCM) are vascular lesions within the central nervous system, consisting of dilated and hemorrhage-prone capillaries. CCMs can cause debilitating neurological symptoms, and surgical excision or stereotactic radiosurgery are the only current treatment options. Meanwhile, transient blood-brain barrier opening (BBBO) with focused ultrasound (FUS) and microbubbles is now understood to exert potentially beneficial bioeffects, such as stimulation of neurogenesis and clearance of amyloid-{beta}. Here, we tested whether FUS BBBO could be deployed therapeutically to control CCM formation and progression in a clinically-representative murine model. METHODSCCMs were induced in mice by postnatal, endothelial-specific Krit1 ablation. FUS was applied for BBBO with fixed peak-negative pressures (PNPs; 0.2-0.6 MPa) or passive cavitation detection-modulated PNPs. Magnetic resonance imaging (MRI) was used to target FUS treatments, evaluate safety, and measure longitudinal changes in CCM growth after BBBO. RESULTSFUS BBBO elicited gadolinium accumulation primarily at the perilesional boundaries of CCMs, rather than lesion cores. Passive cavitation detection and gadolinium contrast enhancement were comparable in CCM and wild-type mice, indicating that Krit1 ablation does not confer differential sensitivity to FUS BBBO. Acutely, CCMs exposed to FUS BBBO remained structurally stable, with no signs of hemorrhage. Longitudinal MRI revealed that FUS BBBO halted the growth of 94% of CCMs treated in the study. At 1 month, FUS BBBO-treated lesions lost, on average, 9% of their pre-sonication volume. In contrast, non-sonicated control lesions grew to 670% of their initial volume. Lesion control with FUS BBBO was accompanied by a marked reduction in the area and mesenchymal appearance of Krit mutant endothelium. Strikingly, in mice receiving multiple BBBO treatments with fixed PNPs, de novo CCM formation was significantly reduced by 81%. Mock treatment plans on MRIs of patients with surgically inaccessible lesions revealed their lesions are amenable to FUS BBBO with current clinical technology. CONCLUSIONSOur results establish FUS BBBO as a novel, non-invasive modality that can safely arrest murine CCM growth and prevent their de novo formation. As an incisionless, MR image-guided therapy with the ability to target eloquent brain locations, FUS BBBO offers an unparalleled potential to revolutionize the therapeutic experience and enhance the accessibility of treatments for CCM patients.

bioengineering↗

bioRxiv2022

Early events marking lung fibroblast transition to profibrotic state in idiopathic pulmonary fibrosis

BackgroundIdiopathic Pulmonary Fibrosis (IPF) is an age-associated progressive lung disease with accumulation of scar tissue impairing gas exchange. Previous high-throughput studies elucidated the role of cellular heterogeneity and molecular pathways in advanced disease. However, critical pathogenic pathways occurring in the transition of fibroblasts from normal to profibrotic have been largely overlooked. MethodsWe used single cell transcriptomics (scRNA-seq) from lungs of healthy controls and IPF patients (lower and upper lobes). We identified fibroblast subclusters, genes and pathways associated to early disease. Immunofluorescence assays validated the role of MOXD1 early in fibrosis. FindingsWe identified four distinct fibroblast subgroups, including one marking the normal-to-profibrotic state transition. Our results show for the first time that global downregulation of ribosomal proteins and significant upregulation of the majority of copper-binding proteins, including MOXD1, mark the IPF transition. We find no significant differences in gene expression in IPF upper and lower lobe samples, which were selected to have low and high degree of fibrosis, respectively. InterpretationEarly events during IPF onset in fibroblasts include dysregulation of ribosomal and copper-binding proteins. Fibroblasts in early stage IPF have already acquired a profibrotic phenotype while hallmarks of advanced disease, including fibroblast foci and honeycomb formation, are still not evident. The new transitional fibroblasts we discover could prove very important for studying the role of fibroblast plasticity in disease progression and help develop early diagnosis tools and therapeutic interventions targeting earlier disease states.

genomics↗

bioRxiv2020

DeepEthogram: a machine learning pipeline for supervised behavior classification from raw pixels

Researchers commonly acquire videos of animal behavior and quantify the prevalence of behaviors of interest to study nervous system function, the effects of gene mutations, and the efficacy of pharmacological therapies. This analysis is typically performed manually and is therefore immensely time consuming, often limited to a small number of behaviors, and variable across researchers. Here, we created DeepEthogram: software that takes raw pixel values of videos as input and uses machine learning to output an ethogram, the set of user-defined behaviors of interest present in each frame of a video. We used convolutional neural network models that compute motion in a video, extract features from motion and single frames, and classify these features into behaviors. These models classified behaviors with greater than 90% accuracy on single frames in videos of flies and mice, matching expert-level human performance. The models accurately predicted even extremely rare behaviors, required little training data, and generalized to new videos and subjects. DeepEthogram runs rapidly on common scientific computer hardware and has a graphical user interface that does not require programming by the end-user. We anticipate DeepEthogram will enable the rapid, automated, and reproducible assignment of behavior labels to every frame of a video, thus accelerating all those studies that quantify behaviors of interest. Code is available at: https://github.com/jbohnslav/deepethogram

neuroscience↗