Human Endothelial Cell Collection from the Middle Cerebral Artery in Acute Ischemic Stroke

Sheth, Sunil A; Verma, Abhishek; Malhotra, Konark; Tang, Xiannan; Tateshima, Satoshi; Jahan, Reza; Hinman, Jason D

doi:10.1016/j.jstrokecerebrovasdis.2017.09.054

Cited by 5 publications

(8 citation statements)

References 8 publications

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“…A possible reason for this could be compromised cell viability due to thrombus effect on the endothelial cells or by the relatively “dirty” cell mix that is obtained in the setting of MT. Complete manual removal of the thrombus from the stent retriever before EC dissociation, as described in the human setting by Sheth et al [16], could possibly resolve this issue.…”

Section: Discussionmentioning

confidence: 99%

“…In this study, we present a pipeline for harvesting, culturing, and then analyzing endothelial cells using FACS and scRNA-seq in a model of mechanical thrombectomy. This pipeline for analysis could readily be integrated with the method of EC harvest in human AIS recently reported by Sheth et al [16]. The time of the procedure is not extended, and the device itself is not affected by the harvesting method.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, an adapted new method of harvesting EC biopsy in AIS was demonstrated [16]. We hypothesized that applicability of this protocol for research would benefit from a molecular biology analysis with next-generation sequencing transcriptomics.…”

Section: Introductionmentioning

confidence: 99%

“…The advent of mechanical thrombectomy to treat acute ischemic stroke (AIS) [ 8 – 12 ] has enabled unprecedented access to thrombotic material [ 13 – 15 ]. Recently, an adapted new method of harvesting EC biopsy in AIS was demonstrated [ 16 ]. We hypothesized that applicability of this protocol for research would benefit from a molecular biology analysis with next-generation sequencing transcriptomics.…”

Section: Introductionmentioning

confidence: 99%

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Transcriptomic analysis of the harvested endothelial cells in a swine model of mechanical thrombectomy

et al. 2018

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PurposeIn mechanical thrombectomy (MT) for ischemic stroke, endothelial cells (ECs) from intracranial blood vessels adhere to the stent retriever device and can be harvested. However, understanding the molecular biology and the role of the endothelium in different pathological conditions remains insufficient. The purpose of the study was to characterize and analyze the molecular aspect of harvested ECs using cell culture and transcriptomic techniques in an MT swine model relevant to clinical ischemic stroke.MethodsIn swine, preformed thrombi were injected into the external carotid and subclavian arteries to occlude their branches. MT was performed according to clinical routine. The stent retriever device and thrombus were treated with cell dissociation buffer. The resulting cell suspension was analyzed by immunohistochemistry and was cultured. Cultured cells were analyzed using single-cell RNA sequencing (scRNA-seq) after fluorescence-activated cell sorting (FACS).ResultsA total number of 37 samples were obtained containing CD31-positive cells. Cell culture was successful in 90% of samples, and the cells expressed multiple typical EC protein markers. Eighty-nine percent of the sorted cells yielded high-quality transcriptomes, and single-cell transcriptomes from cultured cells showed that they expressed typical endothelial gene patterns. Gene expression analysis of ECs from an occluded artery did not show distinctive clustering into subtypes.ConclusionECs harvested during MT can be cultured and analyzed using single-cell transcriptomic techniques. This analysis can be implemented in clinical practice to study the EC gene expression of comorbidities, such as hypertension, diabetes mellitus, and metabolic syndrome, in patients suffering from acute ischemic stroke.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Transcriptomic analysis of the harvested endothelial cells in a swine model of mechanical thrombectomy

et al. 2018

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“…These range from routine analysis of cerebrospinal fluid and brain biopsy samples in selected cases36 to samples uniquely available from patients who suffered a stroke such as histological and proteomic analysis of clots retrieved during thrombectomy37 and cerebrovascular endothelial cells that can be captured from stent retrievers after use in thrombectomy 38. Though presently these additive biological variables are not routinely used, they hold the potential to unlock additional diagnostic, prognostic and theranostic value.…”

Section: Biological Datamentioning

confidence: 99%

Principles of precision medicine in stroke

Hinman

Rost

Leung

et al. 2016

J Neurol Neurosurg Psychiatry

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The era of precision medicine has arrived and conveys tremendous potential, particularly for stroke neurology. The diagnosis of stroke, its underlying aetiology, theranostic strategies, recurrence risk and path to recovery are populated by a series of highly individualised questions. Moreover, the phenotypic complexity of a clinical diagnosis of stroke makes a simple genetic risk assessment only partially informative on an individual basis. The guiding principles of precision medicine in stroke underscore the need to identify, value, organise and analyse the multitude of variables obtained from each individual to generate a precise approach to optimise cerebrovascular health. Existing data may be leveraged with novel technologies, informatics and practical clinical paradigms to apply these principles in stroke and realise the promise of precision medicine. Importantly, precision medicine in stroke will only be realised once efforts to collect, value and synthesise the wealth of data collected in clinical trials and routine care starts. Stroke theranostics, the ultimate vision of synchronising tailored therapeutic strategies based on specific diagnostic data, demand cerebrovascular expertise on big data approaches to clinically relevant paradigms. This review considers such challenges and delineates the principles on a roadmap for rational application of precision medicine to stroke and cerebrovascular health.Precision medicine is broadly defined as tailored diagnosis and therapy for each individual patient.

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Endovascular Device for Endothelial Cell Sampling

Sandell

Chireh

Spyrou

et al. 2022

Advanced NanoBiomed Research

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Endothelial cells play an important role in several vascular diseases, and molecular analysis of these cells could provide valuable information on underlying tissue status. However, no clinically established procedure for harvesting endothelial cells exists. A micro‐3D‐printed device adapted for endovascular techniques to harvest endothelial cells for transcriptomic analysis is presented. In vivo evaluation in swine (n = 6) yielded tissue samples in 60 out of 65 cases, of which 80% show a substantial amount of tissue. The cytological evaluation indicates high selectivity towards endothelial cells, and RNA‐sequencing shows gene expression signatures consistent with vascular tissue. It is found that there are no short‐term safety risks compared to operation with a control wire of equal dimensions and acute complications are not detected. If translated to clinical use, the device could enable increased understanding of early‐stage endothelial cell‐mediated disease progression and earlier diagnosis of diseases such as atherosclerosis.

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Human Endothelial Cell Collection from the Middle Cerebral Artery in Acute Ischemic Stroke

Cited by 5 publications

References 8 publications

Transcriptomic analysis of the harvested endothelial cells in a swine model of mechanical thrombectomy

Transcriptomic analysis of the harvested endothelial cells in a swine model of mechanical thrombectomy

Principles of precision medicine in stroke

Endovascular Device for Endothelial Cell Sampling

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