Background Ischemic stroke research faces difficulties in translating pathology between animal models and human patients to develop treatments. Mechanical thrombectomy, for the first time, offers a momentary window into the changes occurring in ischemia. We developed a tissue banking protocol to capture intracranial thrombi and the blood immediately proximal and distal to it.
Objective To develop and share a reproducible protocol to bank these specimens for future analysis.
Methods We established a protocol approved by the institutional review board for tissue processing during thrombectomy (www.clinicaltrials.gov NCT03153683). The protocol was a joint clinical/basic science effort among multiple laboratories and the NeuroInterventional Radiology service line. We constructed a workspace in the angiography suite, and developed a step-by-step process for specimen retrieval and processing.
Results Our protocol successfully yielded samples for analysis in all but one case. In our preliminary dataset, the process produced adequate amounts of tissue from distal blood, proximal blood, and thrombi for gene expression and proteomics analyses. We describe the tissue banking protocol, and highlight training protocols and mechanics of on-call research staffing. In addition, preliminary integrity analyses demonstrated high-quality yields for RNA and protein.
Conclusions We have developed a novel tissue banking protocol using mechanical thrombectomy to capture thrombus along with arterial blood proximal and distal to it. The protocol provides high-quality specimens, facilitating analysis of the initial molecular response to ischemic stroke in the human condition for the first time. This approach will permit reverse translation to animal models for treatment development.
- inflammatory response
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Contributors JFF is the principal investigator for the BACTRAC tissue bank. Responsibilities included oversight and submission of regulatory documents, design of the tissue bank, collection of tissues, evaluation of data, and drafting of the manuscript. LAC is the administrator of the tissue bank; she catalogues the specimens and tracks their use in experiments. She also played a strong role in design of the banking protocol. AAG also played a strong role in the design of the banking protocol. She oversaw the training of new researchers on the protocol. SRM collected the clinical data, and designed the database for clinical/radiographic data collection and storage. KES, ALT, DNE, and SMD all played roles in consensus-design of the tissue bank. They collected and processed the samples included in the analysis. DEL designed the radiographic imaging data collection methods, and is the neuroradiologist who performs the assessments. AA and SG are neurointerventionalists who perform thrombectomy and collect tissues. JMR oversees the tissue collection training protocol, and assists with tissue collection. GJB and KRP played strong roles in the formation of the interdisciplinary collaboration. In addition, they design questions of the banked tissue, and perform analyses of the tissue for integrity and also for new markers/cytokines.
Funding Initial support for preliminary data was provided through an institutional grant from the University of Kentucky Department of Neurology.
Competing interests None declared.
Patient consent Not required.
Ethics approval University of Kentucky institutional review board.
Provenance and peer review Not commissioned; externally peer reviewed.
Data sharing statement The authors will make the BACTRAC tissue bank collection protocol available upon request. These data are included in the manuscript, but the authors encourage collaboration with other laboratories. Please contact the corresponding author (JFF) for details.