Repeated 5-aminolevulinic acid mediated sonodynamic therapy using magnetic resonance guided focused ultrasound in rat brain tumour models.
Authors: Wu SK, Tsai CL, Mir A, Marcus SL, Hynynen K
Sonodynamic therapy is an emerging therapeutic approach against brain tumours. However, the treatment scheme and ultrasound parameters have yet to be explored for clinical translation. Our study aimed to optimize ultrasound parameters for sonodynamic therapy (SDT) with 5-ALA as a sonosensitizing agent and to evaluate its therapeutic outcome on the rodent 9L gliosarcoma and the human U87 glioblastoma models. We stereotactically implanted brain tumour cells in rats and monitored tumour volume via MRI. SDT was conducted weekly using a 60 mg/kg dose of 5-ALA, injected intravenously 6 h before sonication. We used a driving frequency of 580 kHz with 0.75 MPa and evaluated the effect of different burst lengths to optimize ultrasound parameters. We also tested SDT against advanced-stage brain tumours to verify its efficacy further. Our results showed that a longer burst length could improve therapeutic outcomes. Tumour growth inhibition was established only in the first three weeks with 10 ms and 50 ms burst length sonication, but 86 ms burst length greatly improved the survival outcome. Therefore, the therapeutic efficacy is proportionate to the burst length and, thus, the total delivered energy. Repeated SDT using multiple targets to cover the entire tumour volume with optimal ultrasound parameters can achieve significant anti-tumour effects in both 9L and U87 models. Lastly, our results on late-stage tumour treatments showed that SDT can still provide prolonged survival. These promising findings demonstrate that repeated SDT using transcranial-focused ultrasound together with 5-ALA can optimize anti-tumour effects and even lead to complete clearance of the tumours. This weekly treatment with pulsed ultrasound sonication strategy is practical for future clinical translation.
Introduction
Purpose
Sonodynamic therapy
Study Objective
To optimize ultrasound burst-length parameters for 5-ALA sonodynamic therapy and evaluate its therapeutic efficacy in rodent 9L gliosarcoma and human U87 glioblastoma models.
Animal model / Human subject
Rat (Rattus norvegicus); Strains: Wistar and RNU nude (T-cell deficient); Age: None (weights reported: Wistar 311 ± 22 g, RNU nude 260 ± 24 g); Sex: male
Disease model
Glioma (brain tumours: rodent 9L gliosarcoma and human U87 glioblastoma)
MRI or image guidance method
MRI-guided FUS: preclinical MRI-guided focused ultrasound system (prototype similar to LP100); MRI spatial coordinates co-registered to the spherically focused transducer; contrast-enhanced T1-weighted images used to confirm tumour location and select sonication targets in treatment-planning software.
Targeted brain region(s)
Striatum
Target coordinates
AP +0.5 mm (anterior to bregma), ML +2.8 mm (right lateral to bregma), DV 5.0 mm (depth from dura)
Cargo name and characteristics
5-aminolevulinic acid (5-ALA) — small molecule sonosensitizer; prepared at 60 mg/mL in PBS, administered intravenously (tail vein) at 60 mg/kg dose ~6 hours before focused ultrasound sonication
Route of administration
Intravenous (tail vein injection)
Outcomes and Safety
Summary of Outcomes
Repeated sonodynamic therapy with 5-ALA produced marked anti-tumor effects: in the immunocompetent 9L gliosarcoma model three weekly treatments achieved complete tumor clearance and greatly prolonged survival (including efficacy against late-stage tumors), whereas effects were reduced in the immunodeficient U87 model. Of the ultrasound parameters tested (580 kHz driving frequency, 0.75 MPa, 300 bursts/location, burst lengths 10 ms, 50 ms, 86 ms), only the long 86 ms burst length (with three weekly treatments and 60 mg/kg 5-ALA IV given 6 h prior) was highly effective; 10 ms and 50 ms gave only transient or minimal benefit.
Duration of biological effect
80 days
Safety-related matter
No major adverse effects were reported: animals tolerated SDT well (no body-weight loss), normal brain tissue showed no significant MRI changes, and SDT-treated animals were tumor-free though a slight enlargement of the ventricle on the tumour side was observed. The authors note potential safety concerns with higher pressures or prolonged bursts (skull heating, standing-wave variability, and pre-focal cavitation at the scalp), but state the chosen exposures can be safely delivered through the human skull.
Brain Region
Ultrasound Parameters
Ultrasound instrument
Pre-clinical MRI-guided focused ultrasound system (prototype similar to LP100; FUS Inc., Toronto, ON, Canada); spherically focused transducer: frequency 0.58 MHz, diameter 75 mm, focal number 0.8, focus FWHM 2.9 mm (diameter) × 14 mm (axial)
FUS Frequency
0.58 MHz (580 kHz)
FUS Pressure
0.75 MPa
FUS Mode
pulsed
Pulse duration
10 ms, 50 ms, 86 ms
Duration of a single FUS session
5*N minutes
Focal Characteristics
Focal depth: 14 mm; Focal length: 60 mm (derived from focal number 0.8 and diameter 75 mm); Aperture size: 75 mm
Treatment frequency
multiple
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