Increased PDGFRB and NF-κB signaling caused by highly prevalent somatic mutations in intracranial aneurysms.

Intracranial aneurysms (IAs) are a high-risk factor for life-threatening subarachnoid hemorrhage. Their etiology, however, remains mostly unknown at present. We conducted screening for sporadic somatic mutations in 65 IA tissues (54 saccular and 11 fusiform aneurysms) and paired blood samples by whole-exome and targeted deep sequencing. We identified sporadic mutations in multiple signaling genes and examined their impact on downstream signaling pathways and gene expression in vitro and an arterial dilatation model in mice in vivo. We identified 16 genes that were mutated in at least one IA case and found that these mutations were highly prevalent (92%: 60 of 65 IAs) among all IA cases examined. In particular, mutations in six genes (PDGFRB, AHNAK, OBSCN, RBM10, CACNA1E, and OR5P3), many of which are linked to NF-κB signaling, were found in both fusiform and saccular IAs at a high prevalence (43% of all IA cases examined). We found that mutant PDGFRBs constitutively activated ERK and NF-κB signaling, enhanced cell motility, and induced inflammation-related gene expression in vitro. Spatial transcriptomics also detected similar changes in vessels from patients with IA. Furthermore, virus-mediated overexpression of a mutant PDGFRB induced a fusiform-like dilatation of the basilar artery in mice, which was blocked by systemic administration of the tyrosine kinase inhibitor sunitinib. Collectively, this study reveals a high prevalence of somatic mutations in NF-κB signaling pathway-related genes in both fusiform and saccular IAs and opens a new avenue of research for developing pharmacological interventions.

A preliminary biodistribution study of [99mTc]sodium pertechnetate prepared from an electron linear accelerator and activated carbon-based 99mTc generator.

INTRODUCTION: Production of 99Mo/99mTc using an electron linear accelerator (linac) and activated carbon (AC)-based 99mTc generator (linac-AC) is an alternative approach to the conventional fission production of 99Mo/99mTc. As a preliminary investigation of the clinical applicability of a linac-AC-derived 99mTc radiopharmaceutical, the biodistribution of linac-AC-derived [99mTc]sodium pertechnetate ([99mTc]NaTcO4) was measured and compared against fission-derived [99mTc]NaTcO4 at one time point. METHODS: 99Mo was produced by irradiating nonenriched MoO3 targets with bremsstrahlung photons generated from 55.5-MeV linac electron beams. 99mTc was then separated and purified from the 99Mo using an AC-based 99mTc generator. Subsequently, biodistribution of the linac-AC-derived [99mTc]NaTcO4 in healthy female Slc:ICR mice (n = 6) was measured by dissection and compared with that of fission-derived [99mTc]NaTcO4 (n = 4) at 30 min after injection. RESULTS: The two types of [99mTc]NaTcO4 exhibited similar biodistribution in all the organs and tissues examined: the uptakes of [99mTc]NaTcO4 prepared from the linac-AC method and those prepared from the fission method were 138.9 ± 69.9%ID/g and 160.6 ± 49.2%ID/g in the thyroids, respectively, 33.4 ± 5.5%ID/g and 29.4 ± 9.1%ID/g in the salivary glands, respectively, and less than 10%ID/g in blood and all the other organs. No adverse effects were observed in the mice administered with either [99mTc]NaTcO4. CONCLUSION: The clinical applicability of linac-AC-derived [99mTc]NaTcO4 was suggested by its similar biodistribution with fission-derived [99mTc]NaTcO4 at one time point. Further biodistribution studies at multiple time points are encouraged to demonstrate the bioequivalence between linac-AC- and fission-derived [99mTc]NaTcO4.

Radiolabeling of PSMA-617 with 89Zr: A novel use of DMSO to improve radiochemical yield and preliminary small-animal PET results.

INTRODUCTION: Novel diagnostic and therapeutic options are urgently needed for patients with metastatic castration-resistant prostate cancer (CRPC). PSMA-617 is one of the most promising ligands that bind to prostate specific membrane antigen (PSMA), the cell surface biomarker of CRPC. Of the radiolabeled PSMA ligands developed to date, [68Ga]Ga-PSMA-617 is most commonly used for PSMA positron emission tomography (PET) prior to radioligand therapy (RLT) with [177Lu]Lu-PSMA-617. However, the presence of 68Ga radioactivity (half-life 68 m) in urine at the early PET imaging time point complicates optimization of the therapeutic dose of PSMA-617 labeled with 177Lu (half-life 6.7 d). Thus, PET imaging with the long-lived positron emitter 89Zr (half-life 3.3 d) would be better suited in order to optimize the dose of [177Lu]Lu-PSMA-617 as 89Zr PET allows scans after excretion of the radioactive urine. Until now, PSMA-617 could not be radiolabeled with 89Zr with high radiochemical yield due to poor incorporation of 89Zr into 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA). Here we report a novel method for radiolabeling PSMA-617 with 89Zr and the preliminary results of small-animal PET with [89Zr]Zr-PSMA-617. METHODS: We labeled PSMA-617 with 89Zr in a 1:1 mixture of DMSO and HEPES buffer at 90 °C for 30 min, followed by quality control analysis by HPLC. We then determined the dissociation constant (Kd) and logD values of [89Zr]Zr-PSMA-617. We obtained PET images of [89Zr]Zr-PSMA-617 at 24 h in mice bearing both LNCaP (PSMA-positive) and PC-3 (PSMA-negative) tumors (N = 5). The ex vivo [89Zr]Zr-PSMA-617 biodistribution was then examined separately using tissue samples of LNCaP-bearing mice at 2 h (N = 4) and 24 h (N = 4). RESULTS: [89Zr]Zr-PSMA-617 was prepared with a radiochemical yield of 70 ± 9%. The Kd value was 6.8 ± 3.5 nM. The logD value was -4.05 ± 0.20. PET images showed the highest uptake in LNCaP tumors (maximum standardized uptake value, SUVmax = 0.98 ± 0.32) and low uptake in kidneys (SUVmax = 0.18 ± 0.7) due to the absence of urine radioactivity. CONCLUSION: [89Zr]Zr-PSMA-617 was successfully prepared using DMSO and HEPES buffer. [89Zr]Zr-PSMA-617 visualized PSMA-positive LNCaP tumors in the absence of radioactive urine 24 h p.i. ADVANCES IN KNOWLEDGE AND IMPLICATIONS FOR PATIENT CARE: This method of radiolabeling PSMA-617 with 89Zr using DMSO would be suitable for future clinical trials. Prediction of radiation dose by using [89Zr]Zr-PSMA-617 leads to the safe and effective RLT with [177Lu]Lu-PSMA-617.