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Scanning ultrasound-mediated memory and functional improvements do not require amyloid-β reduction

Gerhard Leinenga, Xuan Vinh To, Liviu-Gabriel Bodea, Jumana Yousef, Gina Richter-Stretton, Tishila Palliyaguru, Antony Chicoteau, Laura Dagley, Fatima Nasrallah, Jürgen Götz

Molecular Psychiatry 2024 · 10.1038/s41380-024-02509-5

rodentalzheimers diseasebehaviourfmriother mrihistology molecular

Abstract

A prevalent view in treating age-dependent disorders including Alzheimer's disease (AD) is that the underlying amyloid plaque pathology must be targeted for cognitive improvements. In contrast, we report here that repeated scanning ultrasound (SUS) treatment at 1 MHz frequency can ameliorate memory deficits in the APP23 mouse model of AD without reducing amyloid-β (Aβ) burden. Different from previous studies that had shown Aβ clearance as a consequence of blood-brain barrier (BBB) opening, here, the BBB was not opened as no microbubbles were used. Quantitative SWATH proteomics and functional magnetic resonance imaging revealed that ultrasound induced long-lasting functional changes that correlate with the improvement in memory. Intriguingly, the treatment was more effective at a higher frequency (1 MHz) than at a frequency within the range currently explored in clinical trials in AD patients (286 kHz). Together, our data suggest frequency-dependent bio-effects of ultrasound and a dissociation of cognitive improvement and Aβ clearance, with important implications for the design of trials for AD therapies.

Abstract via europepmc.

Speciesmouse
Subjects12, 11swept animals
Sessions per subject8
Randomisedno
Blindingnot reported
Sham / controlinactive transducer
Auditory controlnot reported
Readout timingoffline
Anaesthesiaanaesthetised
Readoutsbehaviour, fmri, other mri, histology molecularActive place avoidance spatial memory test; resting-state and stimulus-evoked fMRI; T2-weighted structural MRI and NODDI diffusion MRI; SWATH-MS quantitative proteomics; amyloid-beta ELISA; Campbell-Switzer silver staining for plaques; Western blot for synaptic tau
Direction of effectexcitatoryHighF (1 MHz) scanning ultrasound without microbubbles improved spatial memory (fewer shocks, better learning index) in APP23 mice relative to sham, increased resting-state functional connectivity in memory-related networks, and altered the cortical proteome, all without reducing amyloid-beta plaque burden; LowF (286 kHz) produced smaller, largely non-significant behavioural improvements.
Adverse eventsnot reported

Exposures

Exposure 1: HighF: 1 MHz scanning ultrasound (SUS-only) to whole brain

Target: whole brain or unfocused — “whole brain (5x6 grid of scanning spots)
Device: Philips Research · Philips Research · Therapy Imaging Probe System (TIPS), 8-element annular array

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)1,000✓✓
Pulse duration (ms)10✓✓
Pulse repetition frequency (Hz)10✓✓
Duty cycle (%)not reportedpulse duration × PRF gives 10%
Sonication duration (s)6✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)700✓✓
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)not reported
In-situ estimatederatingsingle value
In-situ pressure (kPa)570✓✓
In-situ Isppa (W/cm²)not reported
In-situ Ispta (W/cm²)not reported
Protocol, in the paper’s words

Whole-brain exposure achieved by scanning a 5x6 grid of spots (1.5 mm spacing) with the annular-array TIPS transducer; mice received 8 once-per-week treatments; sham mice received anaesthesia and transducer placement without power.

Exposure 2: LowF: 286 kHz scanning ultrasound (SUS-only) to whole brain

Target: whole brain or unfocused — “whole brain (4-spot grid)
Device: Sonic Concepts · Sonic Concepts, Bothell WA, USA · H-117

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)286✓✓
Pulse duration (ms)10✓✓
Pulse repetition frequency (Hz)10✓✓
Duty cycle (%)not reportedpulse duration × PRF gives 10%
Sonication duration (s)6✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)not reported
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)not reported
In-situ estimatederatingsingle value
In-situ pressure (kPa)570✓✓
In-situ Isppa (W/cm²)not reported
In-situ Ispta (W/cm²)not reported
Protocol, in the paper’s words

Whole-brain exposure achieved with a 4-spot grid (6 mm spacing) using the H-117 transducer; mice received 8 once-per-week treatments; sham mice received anaesthesia and transducer placement without power.

Flags from extraction

  • n_subjectsOnly per-group sizes are given for the two ultrasound-treated groups (HighF N=12, LowF N=11); no combined total exposed to ultrasound is stated, so group sizes are listed rather than summed. The sham (N=12) and WT (N=12) groups were not exposed to real ultrasound and are excluded.
  • exposures[1].free_field.pressure_kpaFor the LowF condition the paper calls the water-measured 0.57 MPa value 'in situ' (assuming negligible skull attenuation) rather than free-field, so it was placed in in_situ rather than free_field despite being measured in water.