Hamza Farooq

Intertumoral Heterogeneity within Medulloblastoma Subgroups

While molecular subgrouping has revolutionized medulloblastoma classification, the extent of heterogeneity within subgroups is unknown. Similarity network fusion (SNF) applied to genome-wide DNA methylation and gene expression data across 763 primary samples identifies very homogeneous clusters of patients, supporting the presence of medulloblastoma subtypes. After integration of somatic copy-number alterations, and clinical features specific to each cluster, we identify 12 different subtypes of medulloblastoma. Integrative analysis using SNF further delineates group 3 from group 4 medulloblastoma, which is not as readily apparent through analyses of individual data types. Two clear subtypes of infants with Sonic Hedgehog medulloblastoma with disparate outcomes and biology are identified. Medulloblastoma subtypes identified through integrative clustering have important implications for stratification of future clinical trials.

Spatial heterogeneity in medulloblastoma

Spatial heterogeneity of transcriptional and genetic markers between physically isolated biopsies of a single tumor poses major barriers to the identification of biomarkers and the development of targeted therapies that will be effective against the entire tumor. We analyzed the spatial heterogeneity of multiregional biopsies from 35 patients, using a combination of transcriptomic and genomic profiles. Medulloblastomas (MBs), but not high-grade gliomas (HGGs), demonstrated spatially homogeneous transcriptomes, which allowed for accurate subgrouping of tumors from a single biopsy. Conversely, somatic mutations that affect genes suitable for targeted therapeutics demonstrated high levels of spatial heterogeneity in MB, malignant glioma, and renal cell carcinoma (RCC). Actionable targets found in a single MB biopsy were seldom clonal across the entire tumor, which brings the efficacy of monotherapies against a single target into question. Clinical trials of targeted therapies for MB should first ensure the spatially ubiquitous nature of the target mutation.

Evaluation of flow velocities after carotid artery stenting through split spectrum Doppler optical coherence tomography and computational fluid dynamics modeling

Hemodynamics plays a critical role in the development of atherosclerosis, specifically in regions of curved vasculature such as bifurcations exhibiting irregular blood flow profiles. Carotid atherosclerotic disease can be intervened by stent implantation, but this may result in greater alterations to local blood flow and consequently further complications. This study demonstrates the use of a variant of Doppler optical coherence tomography (DOCT) known as split spectrum DOCT (ssDOCT) to evaluate hemodynamic patterns both before and after stent implantation in the bifurcation junction in the internal carotid artery (ICA). Computational fluid dynamics (CFD) models were constructed to simulate blood velocity profiles and compared to the findings achieved through ssDOCT images. Both methods demonstrated noticeable alterations in hemodynamic patterns following stent implantation, with features such as slow velocity regions at the neck of the bifurcation and recirculation zones at the stent struts. Strong correlation between CFD models and ssDOCT images demonstrate the potential of ssDOCT imaging in the optimization of stent implantation in the clinical setting.

High-resolution imaging of the central nervous system: how novel imaging methods combined with navigation strategies will advance patient care.

This narrative review captures a subset of recent advances in imaging of the central nervous system. First, we focus on improvements in the spatial and temporal profile afforded by optical coherence tomography, fluorescence-guided surgery, and Coherent Anti-Stokes Raman Scattering Microscopy. Next, we highlight advances in the generation and uses of imaging-based atlases and discuss how this will be applied to specific clinical situations. To conclude, we discuss how these and other imaging tools will be combined with neuronavigation techniques to guide surgeons in the operating room. Collectively, this work aims to highlight emerging biomedical imaging strategies that hold potential to be a valuable tool for both clinicians and researchers in the years to come.

Childhood cerebellar tumors mirror conserved fetal transcriptional programs

The study of the origin and development of cerebellar tumours has been hampered by the complexity and heterogeneity of cerebellar cells that change over the course of development. We used single-cell transcriptomics to study >60,000 cells from the developing murine cerebellum, and show that different molecular subgroups of childhood cerebellar tumors mirror the transcription of cells from distinct, temporally restricted cerebellar lineages. Sonic Hedgehog medulloblastoma transcriptionally mirrors the granule cell hierarchy as expected, whereas Group 3 medulloblastoma resemble Nestin+ve stem cells, Group 4 medulloblastomas resemble unipolar brush cells, and PFA/PFB ependymoma and cerebellar pilocytic astrocytoma resemble the prenatal gliogenic progenitor cells. Furthermore, single-cell transcriptomics of human childhood cerebellar tumors demonstrates that many bulk tumors contain a mixed population of cells with divergent differentiation. Our data highlight cerebellar tumors as a disorder of early brain development, and provide a proximate explanation for the peak incidence of cerebellar tumors in early childhood.

High Speed, High Density Intraoperative 3D Optical Topographical Imaging with Efficient Registration to MRI and CT for Craniospinal Surgical Navigation

Intraoperative image-guided surgical navigation for craniospinal procedures has significantly improved accuracy by providing an avenue for the surgeon to visualize underlying internal structures corresponding to the exposed surface anatomy. Despite the obvious benefits of surgical navigation, surgeon adoption remains relatively low due to long setup and registration times, steep learning curves, and workflow disruptions. We introduce an experimental navigation system utilizing optical topographical imaging (OTI) to acquire the 3D surface anatomy of the surgical cavity, enabling visualization of internal structures relative to exposed surface anatomy from registered preoperative images. Our OTI approach includes near instantaneous and accurate optical measurement of >250,000 surface points, computed at >52,000 points-per-second for considerably faster patient registration than commercially available benchmark systems without compromising spatial accuracy. Our experience of 171 human craniospinal surgical procedures, demonstrated significant workflow improvement (41 s vs. 258 s and 794 s, p < 0.05) relative to benchmark navigation systems without compromising surgical accuracy. Our advancements provide the cornerstone for widespread adoption of image guidance technologies for faster and safer surgeries without intraoperative CT or MRI scans. This work represents a major workflow improvement for navigated craniospinal procedures with possible extension to other image-guided applications.

BMI1 is a therapeutic target in recurrent medulloblastoma

Medulloblastoma (MB) is the most frequent malignant pediatric brain tumor, representing 20% of newly diagnosed childhood central nervous system malignancies. Although advances in multimodal therapy yielded a 5-year survivorship of 80%, MB still accounts for the leading cause of childhood cancer mortality. In this work, we describe the epigenetic regulator BMI1 as a novel therapeutic target for the treatment of recurrent human Group 3 MB, a childhood brain tumor for which there is virtually no treatment option beyond palliation. Current clinical trials for recurrent MB patients based on genomic profiles of primary, treatment-naive tumors will provide limited clinical benefit since recurrent metastatic MBs are highly genetically divergent from their primary tumor. Using a small molecule inhibitor against BMI1, PTC-028, we were able to demonstrate complete ablation of self-renewal of MB stem cells in vitro. When administered to mice xenografted with patient tumors, we observed significant reduction in tumor burden in both local and metastatic compartments and subsequent increased survival, without neurotoxicity. Strikingly, serial in vivo re-transplantation assays demonstrated a marked reduction in tumor initiation ability of recurrent MB cells upon re-transplantation of PTC-028-treated cells into secondary recipient mouse brains. As Group 3 MB is often metastatic and uniformly fatal at recurrence, with no current or planned trials of targeted therapy, an efficacious targeted agent would be rapidly transitioned to clinical trials.

Convergence of BMI1 and CHD7 on ERK Signaling in Medulloblastoma

Summary We describe molecular convergence between BMI1 and CHD7 in the initiation of medulloblastoma. Identified in a functional genomic screen in mouse models, a BMI1High;CHD7Low expression signature within medulloblastoma characterizes patients with poor overall survival. We show that BMI1-mediated repression of the ERK1/2 pathway leads to increased proliferation and tumor burden in primary human MB cells and in a xenograft model, respectively. We provide evidence that repression of the ERK inhibitor DUSP4 by BMI1 is dependent on a more accessible chromatin configuration in G4 MB cells with low CHD7 expression. These findings extend current knowledge of the role of BMI1 and CHD7 in medulloblastoma pathogenesis, and they raise the possibility that pharmacological targeting of BMI1 or ERK may be particularly indicated in a subgroup of MB with low expression levels of CHD7.

Accuracy of image-guided surgical navigation using near infrared (NIR) optical tracking

Spinal surgery is particularly challenging for surgeons, requiring a high level of expertise and precision without being able to see beyond the surface of the bone. Accurate insertion of pedicle screws is critical considering perforation of the pedicle can result in profound clinical consequences including spinal cord, nerve root, arterial injury, neurological deficits, chronic pain, and/or failed back syndrome. Various navigation systems have been designed to guide pedicle screw fixation. Computed tomography (CT)-based image guided navigation systems increase the accuracy of screw placement allowing for 3- dimensional visualization of the spinal anatomy. Current localization techniques require extensive preparation and introduce spatial deviations. Use of near infrared (NIR) optical tracking allows for realtime navigation of the surgery by utilizing spectral domain multiplexing of light, greatly enhancing the surgeon’s situation awareness in the operating room. While the incidence of pedicle screw perforation and complications have been significantly reduced with the introduction of modern navigational technologies, some error exists. Several parameters have been suggested including fiducial localization and registration error, target registration error, and angular deviation. However, many of these techniques quantify error using the pre-operative CT and an intra-operative screenshot without assessing the true screw trajectory. In this study we quantified in-vivo error by comparing the true screw trajectory to the intra-operative trajectory. Pre- and post- operative CT as well as intra-operative screenshots were obtained for a cohort of patients undergoing spinal surgery. We quantified entry point error and angular deviation in the axial and sagittal planes.

Evaluation of hemodynamics changes during interventional stent placement using Doppler optical coherence tomography

Carotid atherosclerosis is a critical medical concern that can lead to ischemic stroke. Local hemodynamic patterns have also been associated with the development of atherosclerosis, particularly in regions with disturbed flow patterns such as bifurcations. Traditionally, this disease was treated using carotid endarterectomy, however recently there is an increasing trend of carotid artery stenting due to its minimally invasive nature. It is well known that this interventional technique creates changes in vasculature geometry and hemodynamic patterns due to the interaction of stent struts with arterial lumen, and is associated with complications such as distal emboli and restenosis. Currently, there is no standard imaging technique to evaluate regional hemodynamic patterns found in stented vessels. Doppler optical coherence tomography (DOCT) provides an opportunity to identify in vivo hemodynamic changes in vasculature using high-resolution imaging. In this study, blood flow profiles were examined at the bifurcation junction in the internal carotid artery (ICA) in a porcine model following stent deployment. Doppler imaging was further conducted using pulsatile flow in a phantom model, and then compared to computational fluid dynamics (CFD) simulation of a virtual bifurcation to assist with the interpretation of emphin vivo results.