Abstract Gadolinium (Gd) enhanced MRI is nonspecific for identifying aggressive glioblastoma biology. Glioblastoma-associated macrophages (GAMs) are pivotal in regrowth. Segregation and Extravascular Localization of Ferumoxytol Imaging (SELFI) is a quantitative immune specific MRI technique. SELFI+ values quantify M2 GAMs by phagocytized Ferumoxytol (Fe), while SELFI- values delineate intravascular Fe, cerebral blood volume. The relationship between SELFI and biology driving therapeutic resistance remains unknown. We aimed to determine if SELFI identified immunosuppressive GAMs and biology contributing to poor outcomes, a capability not yet achievable in clinical practice. This prospective study included four patients with glioblastoma. All underwent preoperative Gd and Fe (7 mg/kg up to 510 mg) enhanced MRI allowing for image guided biopsy based on Gd enhancement and FLAIR hyperintensity. RNA was analyzed on GeneChip Clariom Arrays. Intraoperative biopsy location was referenced to MRI for placement of volumes of interest (VOIs). VOIs quantified MRI phenotype and SELFI values. Gene Set Enrichment Analysis (GSEA) assessed differential expression between MRI features. T-test assessed differences in SELFI (p <0.05) and GSEA significance was assessed by normalized enrichment score (NES) and false discovery rate (FDR) q-value. Four patients provided 36 image guided tissue samples with sufficient quality to assess the causal relationships between MRI features and transcriptomics. Stratification by patient MGMT methylation status (N= 3 unmethylated) demonstrated Gd+ regions had significantly elevated SELFI+ values (Mean±SD, Gd+ = 0.24±0.17, Gd- = 0.08±0.09, P<0.01). GSEA demonstrated that tissues with elevated SELFI+ and lowered SELFI- values were comprised of significantly upregulated hypoxia and transforming growth factor-beta (TGFβ) signaling; among others (NES>1.5, FDR q-value<0.05). SELFI MRI uniquely quantifies glioblastoma by localizing sites of biological aggressiveness known to contribute to therapeutic resistance and disease recurrence. Patient enrollment continues to further define the causal relationships between SELFI, GAMs, and glioblastoma biology.
