Breakthrough in Mechanobiology: NUS Team Pinpoints Piezo1's Force Activation

Piezo1, a key mechanosensitive ion channel, plays a crucial role in how cells sense and respond to mechanical forces, from blood flow regulation to immune responses. Researchers at the National University of Singapore (NUS) have achieved a milestone by directly quantifying the precise force required to activate this channel, clocking in at approximately 15 piconewtons (pN). Published in the prestigious journal Nature Sensors on April 3, 2026, this study marks a significant advancement in understanding mechanotransduction—the process by which cells convert physical stimuli into biochemical signals.

Led by Principal Investigator Jie Yan from the Mechanobiology Institute and Department of Physics at NUS, the team—including Mingyu Sui, Jingzhun Liu, Chaoyu Fu, Yuxia Liu, and Xiaogang Liu from NUS Chemistry and the N.1 Institute for Health—developed an innovative DNA-tethered extracellular force sensing platform. This method bypasses traditional challenges where applied forces were confounded by membrane deformation, allowing for the first direct measurement of Piezo1's activation threshold.

Understanding Piezo1: The Mechanosensitive Powerhouse

Piezo1 (full name: Piezo-type mechanosensitive ion channel component 1) is a trimeric protein forming a large ion channel in cell membranes. Discovered in 2010, it opens in response to mechanical stress, permitting calcium ions to flood into the cell and trigger downstream signaling. This channel is vital for vascular development, red blood cell volume regulation, and even bone formation. Dysfunctions in Piezo1 are linked to conditions like xerocytosis (a hemolytic anemia) and distal arterial aneurysms, highlighting its medical relevance.

In Singapore's thriving biomedical research landscape, NUS has positioned itself at the forefront of mechanobiology studies. The Mechanobiology Institute (MBI), established in 2009, fosters interdisciplinary work combining physics, chemistry, and biology to decode how forces shape life processes. This Piezo1 study exemplifies NUS's commitment to high-impact research, supported by funding from Singapore's National Research Foundation and Ministry of Education.

The Challenge: Measuring Force Without Membrane Interference

Prior methods to study Piezo1 activation relied on techniques like patch-clamp electrophysiology or cell poking, which inevitably deformed the lipid bilayer. This coupling made it impossible to isolate the pure force threshold for channel gating. Researchers hypothesized two main models: a 'force-from-lipids' mechanism (where membrane tension directly opens the channel) versus a 'force-from-filament' model (where cytoskeletal tethers pull on the channel).

The NUS team addressed this by engineering cells expressing a fusion protein: Piezo1 linked to jGCaMP8m, a genetically encoded calcium indicator. This allowed real-time visualization of channel opening via fluorescence changes as calcium enters.

Innovative DNA-Tethered Platform: Precision at Piconewton Scale

The core innovation is a DNA-based tether system using lambda (λ) DNA modified with handles, connected to a micropipette-held bead coated in DSPE-PEG-biotin for cell attachment. Forces are calibrated using DNA hairpins with known rupture forces (e.g., 1000 bp hairpin), verified via magnetic tweezers.

In experiments, researchers used micropipette manipulation in force-jump and force-ramp modes. When tether forces exceeded ~15 pN, calcium transients were observed, even in cells treated with GsMTx4 (a Piezo1 inhibitor that doesn't block tether-pulling). This confirmed activation independent of membrane tension, favoring the force-from-filament model.

The platform's versatility shines: it delivers calibrated piconewton forces directly to extracellular domains while monitoring activity optically, sidestepping electrical noise in traditional setups.

Key Findings: 15 pN Threshold and Membrane Independence

The study revealed Piezo1 activates at approximately 15.0 pN per tether, with rapid response times. Force-ramp experiments showed a clear threshold, beyond which channel opening probability surged. Critically, 'bead-grab' assays—pulling beads without suction—still elicited responses, proving no membrane deformation needed.

Quote from the paper: "Piezo1 opens approximately at 15.0 pN, providing a direct quantification of its activation threshold and demonstrating that Piezo1 can be gated by tether-mediated forces independent of membrane tension, supporting a force from filament mechanism."

Photo by National Cancer Institute on Unsplash

• Threshold force: ~15 pN
• Independent of membrane tension
• Supports force-from-filament model
• Compatible with live-cell calcium imaging

Technical Mastery: From DNA Calibration to Single-Molecule Precision

Calibration involved synthesizing DNA hairpins and using magnetic tweezers to map force-extension curves. Micropipette suction forces matched hairpin rupture events, ensuring accuracy. Cells were HEK293T expressing Piezo1-jGCaMP8m, validated with ionomycin (calcium ionophore) controls showing robust fluorescence.

This step-by-step process—DNA tether attachment, bead grabbing, force application, and ratiometric imaging—provides a blueprint for studying other mechanosensors like Piezo2 or integrins.

Implications for Medicine and Beyond

Knowing Piezo1's exact activation force opens doors to targeted therapies. In vascular diseases, modulating this threshold could prevent excessive channel opening leading to aneurysms. In immunity, it could enhance T-cell mechanosensing for better cancer immunotherapies. For drug discovery, high-throughput screening of Piezo1 agonists/antagonists becomes feasible with calibrated forces.

The platform extends to materials science, testing synthetic mechanosensors or nanomaterials under biological forces. In Singapore, where biotech investments exceed SGD 25 billion via the Research, Innovation and Enterprise 2025 plan, such tools accelerate translation from lab to clinic.Read the full study in Nature Sensors

NUS's Mechanobiology Leadership in Singapore

NUS's Mechanobiology Institute, directed by luminaries like Yan Jie, integrates physics and biology to tackle force-related diseases. Collaborations with N.1 Institute for Health and Tianjin University underscore Singapore's global research hub status. This study, funded by Singapore agencies, boosts NUS's QS ranking (top 10 Asia-wide) and attracts talent to its PhD programs in biophysics.

Singapore universities like NTU and SMU complement this with AI-mechanobiology fusions, but NUS leads in single-molecule force spectroscopy.

Stakeholder Perspectives and Broader Context

Experts hail the work: Reviewers noted its "generalizable strategy for precisely probing mechanotransduction pathways." Jie Yan's prior studies on filopodia forces laid groundwork, showing NUS's sustained excellence.

In Singapore's higher education, where research output grew 15% yearly, this publication elevates mechanobiology, inspiring students via MBI's training programs.

Future Outlook: Scaling to Complex Systems

Next steps include multiplexing for multiple channels, in vivo applications via optogenetics, and AI integration for force prediction. Potential: Designer drugs tuning Piezo1 sensitivity for hypertension or inflammation. NUS plans open-sourcing the platform, fostering collaborations.

As Singapore invests SGD 22 billion in RIE 2025, expect more such breakthroughs, positioning local unis as mechanobiology leaders.

Photo by National Cancer Institute on Unsplash

Actionable Insights for Researchers and Students

• Replicate with commercial λ-DNA kits and jGCaMP8m plasmids.
• Explore variants like disease-linked Piezo1 mutants (e.g., R245H).
• Apply to Singapore fellowships at MBI for mechanobiology PhDs.

This NUS study not only decodes a fundamental biological switch but empowers the next generation of Singaporean scientists.