Single cell transcriptomic analysis of the tumor microenvironment in prostate cancer spine metastasis and an adjacent normal vertebral body

Sunday, May 5, 2024

2:15 PM – 2:17 PM CT

Location: Booth 745 Neuro Spotlight

Presenting Author(s)

Eli Johnson, MD

Resident Physician
Department of Neurosurgery, Duke University School of Medicine, Durham, NC
Durham, NC, US

Co-Author(s)

CB

César D. Baëta, n/a

Clinical Research Assistant
Duke University Hospital, United States
Rory Goodwin, MD PhD (he/him/his)

Associate Professor
Department of Neurosurgery, Duke University
Durham, North Carolina, United States
CH

Corinne Haines, PhD

Postdoctoral Associate
Duke University Hospital, United States
Harrison A. Hockenberry, BS

Senior Clinical Research Specialist
Department of Neurosurgery, Duke University School of Medicine, Durham, NC, United States
Eli Johnson, MD

Resident Physician
Department of Neurosurgery, Duke University School of Medicine, Durham, NC
Durham, NC, US

Submitter(s)

Eli Johnson, MD

Resident Physician
Department of Neurosurgery, Duke University School of Medicine, Durham, NC
Durham, NC, US

Co-Author(s)

Annee D. Nguyen, MS (she/they pronouns)

Basic Science Lab Manager
Department of Neurosurgery, Duke University School of Medicine, Durham, NC
Durham, North Carolina, United States
MP

Meghan J. Price, MD

Resident
Johns Hopkins Medicine
Durham, North Carolina, United States

Disclosure(s):

Eli Johnson, MD: No financial relationships to disclose

Introduction: Spine metastases can result in debilitating complications, such as sensorimotor dysfunction, paralysis, and spinal instability, and account for 90% of spine tumor cases. With few prognostication markers of disease, limited eligibility of spine tumor patients to participate in clinical trials, and thus a paucity of therapeutic targets, there is a distinct need for molecular profiling of spine tumors. To demonstrate the ability to compare the genetic signatures and microenvironments of difficult-to-process osseous metastases and adjacent normal vertebral tissue, we describe an OR-to-bench-top processing of spine tumors for single-cell omic studies, with samples from a metastatic prostate cancer patient.

Methods: Patients undergoing spine surgery with the Department of Neurosurgery at Duke University were consented for tissue collection in accordance with IRB Protocol #Pro00101198. Osseous metastases and adjacent normal vertebral tissue were confirmed histologically (H&E, immunohistochemistry) and radiologically (MRI, PET, CT, and x-ray). Samples were manually dissociated and enzymatically homogenized with collagenase A and DNAse I. Cells were filter sterilized, erythrolyzed, counted to determine live-death ratio, resuspended to 10-20 million cells per mL, then cryopreserved for downstream standard library preparation for single-cell sequencing. Downstream analysis was performed using Seurat pipeline, InferCNV analysis, and SingleR.

Results: Radiological imaging and H&E staining confirmed the presence of T7 osseous metastasis, and P404S and PSA expression confirmed tumor was prostate in origin. Uniform manifold approximation and project (UMAP) plots identified 12 unique clusters of immune cell subpopulations between normal and tumorous vertebrae, with their own subsets of differentially expressed genes.

Conclusion : Processing of spinal vertebral metastases is feasible and can generate granular single cell molecular profiling for spine tumors. Applications of this protocol can aid in identifying therapeutic targets, molecular predictors of disease, and potentiate increased eligibility of patients for clinical trials.